Methods of modulating bone remodeling

ABSTRACT

Disclosed herein, in certain instances, are methods of inhibiting osteoclast differentiation, bone resorption, bone formation, and bone remodeling in an individual in need thereof, comprising administering to the individual a composition comprising substantially fetal support tissue product including amniotic membrane and umbilical cord or an extract thereof, or a composition comprising substantially isolated HC-HA complex.

CROSS-REFERENCE

This application claims priority to International Application No.PCT/US2012/035678 filed on Apr. 27, 2012, which claims priority to U.S.Provisional Application No. 61/480,281, filed Apr. 28, 2011, both ofwhich are incorporated herein in by reference in their entireties.

SUMMARY OF THE INVENTION

Disclosed herein, in certain embodiments, are methods of inhibiting boneresorption in an individual in need thereof, comprising administering tothe individual a composition comprising fetal support tissue product oran extract thereof. In some embodiments, the fetal support tissueproduct comprises placental amniotic membrane (PAM), umbilical cordamniotic membrane (UCAM), placenta, umbilical cord, chorion,amnion-chorion, or any combinations thereof. In some embodiments, thefetal support tissue product comprises frozen or previously frozenplacental amniotic membrane (PAM), frozen or previously frozen umbilicalcord amniotic membrane (UCAM), frozen or previously frozen placenta,frozen or previously frozen umbilical cord, frozen or previously frozenchorion, frozen or previously frozen amnion-chorion, or any combinationsthereof. In some embodiments, the fetal support tissue product in theform of a pulverized powder or a homogenate. In some embodiments, thefetal support tissue product is cryopreserved, lyophilized, terminallysterilized, or a combination thereof. In some embodiments, the extractcomprises HC-HA. In some embodiments, the extract is prepared byultracentrifugation. In some embodiments, the extract is prepared by atleast 2 rounds of ultracentrifugation. In some embodiments, the extractis prepared by more than 2 rounds of untracentrifugation. In someembodiments, the extract is prepared by at least 4 rounds ofuntracentrifugation. In some embodiments, the composition furthercomprises a pharmaceutically-acceptable diluent, excipient or carrier.In some embodiments, the composition is formulated as a solution,suspension, paste, ointment, oil, emulsion, microemulsion, cream,lotion, gel, or combination thereof. In some embodiments, thecomposition is formulated for controlled, sustained, or delayed release.In some embodiments, the composition is injected into an osteolyticjoint. In some embodiments, the composition is administered by a patch.In some embodiments, the patch is placed on an osteolytic bone or anosteolytic joint. In some embodiments, the composition is administeredby an implant. In some embodiments, the implant is placed on anosteolytic bone or an osteolytic joint. In some embodiments, thecomposition is administered by an orthopaedic prosthesis. In someembodiments, the composition is administered by a bone stent. In someembodiments, the composition is a sheet and is wrapped around anosteolytic bone or an osteolytic joint. In some embodiments, the methodsfurther comprise administering to the individual a calcium supplement.

Disclosed herein, in certain embodiments, are methods of treating adisease, disorder, or condition characterized by excessive or undesiredbone resorption, the method comprising administering to an individual inneed thereof a composition comprising fetal support tissue product or anextract thereof. In some embodiments, the fetal support tissue productcomprises placental amniotic membrane (PAM), umbilical cord amnioticmembrane (UCAM), placenta, umbilical cord, chorion, amnion-chorion, orany combinations thereof. In some embodiments, the fetal support tissueproduct comprises frozen or previously frozen placental amnioticmembrane (PAM), frozen or previously frozen umbilical cord amnioticmembrane (UCAM), frozen or previously frozen placenta, frozen orpreviously frozen umbilical cord, frozen or previously frozen chorion,frozen or previously frozen amnion-chorion, or any combinations thereof.In some embodiments, the fetal support tissue product in the form of apulverized powder or a homogenate. In some embodiments, the fetalsupport tissue product is cryopreserved, lyophilized, terminallysterilized, or a combination thereof. In some embodiments, the extractcomprises HC-HA. In some embodiments, the extract is prepared byultracentrifugation. In some embodiments, the extract is prepared by atleast 2 rounds of ultracentrifugation. In some embodiments, the extractis prepared by more than 2 rounds of untracentrifugation. In someembodiments, the extract is prepared by at least 4 rounds ofuntracentrifugation. In some embodiments, the composition furthercomprises a pharmaceutically-acceptable diluent, excipient or carrier.In some embodiments, the composition is formulated as a solution,suspension, paste, ointment, oil, emulsion, microemulsion, cream,lotion, gel, or combination thereof. In some embodiments, thecomposition is formulated for controlled, sustained, or delayed release.In some embodiments, the composition is injected into an osteolyticjoint. In some embodiments, the composition is administered by a patch.In some embodiments, the patch is placed on an osteolytic bone or anosteolytic joint. In some embodiments, the composition is administeredby an implant. In some embodiments, the implant is placed on anosteolytic bone or an osteolytic joint. In some embodiments, thecomposition is administered by an orthopaedic prosthesis. In someembodiments, the composition is administered by a bone stent. In someembodiments, the composition is a sheet and is wrapped around anosteolytic bone or an osteolytic joint. In some embodiments, the methodsfurther comprise administering to the individual a calcium supplement.In some embodiments, the disease, disorder, or condition characterizedby excessive or undesired osteoclast differentiation is arthritis,osteoporosis, a bone tumor, Paget's Disease, alveolar bone degradation,or any combination thereof. In some embodiments, the arthritis isosteoarthritis, rheumatoid arthritis, psoriatic arthritis, or anycombination thereof.

Disclosed herein, in certain embodiments, are methods of promoting orinducing bone formation in an individual in need thereof in anindividual in need thereof, comprising administering to the individual acomposition comprising fetal support tissue product or an extractthereof. In some embodiments, the fetal support tissue product comprisesplacental amniotic membrane (PAM), umbilical cord amniotic membrane(UCAM), placenta, umbilical cord, chorion, amnion-chorion, or anycombinations thereof. In some embodiments, the fetal support tissueproduct comprises frozen or previously frozen placental amnioticmembrane (PAM), frozen or previously frozen umbilical cord amnioticmembrane (UCAM), frozen or previously frozen placenta, frozen orpreviously frozen umbilical cord, frozen or previously frozen chorion,frozen or previously frozen amnion-chorion, or any combinations thereof.In some embodiments, the fetal support tissue product in the form of apulverized powder or a homogenate. In some embodiments, the fetalsupport tissue product is cryopreserved, lyophilized, terminallysterilized, or a combination thereof. In some embodiments, the extractcomprises HC-HA. In some embodiments, the extract is prepared byultracentrifugation. In some embodiments, the extract is prepared by atleast 2 rounds of ultracentrifugation. In some embodiments, the extractis prepared by more than 2 rounds of untracentrifugation. In someembodiments, the extract is prepared by at least 4 rounds ofuntracentrifugation. In some embodiments, the composition furthercomprises a pharmaceutically-acceptable diluent, excipient or carrier.In some embodiments, the composition is formulated as a solution,suspension, paste, ointment, oil, emulsion, microemulsion, cream,lotion, gel, or combination thereof. In some embodiments, thecomposition is formulated for controlled, sustained, or delayed release.In some embodiments, the composition is injected into an osteolyticjoint. In some embodiments, the composition is administered by a patch.In some embodiments, the patch is placed on an osteolytic bone or anosteolytic joint. In some embodiments, the composition is administeredby an implant. In some embodiments, the implant is placed on anosteolytic bone or an osteolytic joint. In some embodiments, thecomposition is administered by an orthopaedic prosthesis. In someembodiments, the composition is administered by a bone stent. In someembodiments, the composition is a sheet and is wrapped around anosteolytic bone or an osteolytic joint. In some embodiments, the methodsfurther comprise administering to the individual a calcium supplement.

Disclosed herein, in certain embodiments, are methods of treating adisease, disorder or condition characterized by a deficiency of boneformation in an individual in need thereof, the method comprisingadministering to an individual in need thereof a composition comprisingfetal support tissue product or an extract thereof. In some embodiments,the fetal support tissue product comprises placental amniotic membrane(PAM), umbilical cord amniotic membrane (UCAM), placenta, umbilicalcord, chorion, amnion-chorion, or any combinations thereof. In someembodiments, the fetal support tissue product comprises frozen orpreviously frozen placental amniotic membrane (PAM), frozen orpreviously frozen umbilical cord amniotic membrane (UCAM), frozen orpreviously frozen placenta, frozen or previously frozen umbilical cord,frozen or previously frozen chorion, frozen or previously frozenamnion-chorion, or any combinations thereof. In some embodiments, thefetal support tissue product in the form of a pulverized powder or ahomogenate. In some embodiments, the fetal support tissue product iscryopreserved, lyophilized, terminally sterilized, or a combinationthereof. In some embodiments, the extract comprises HC-HA. In someembodiments, the extract is prepared by ultracentrifugation. In someembodiments, the extract is prepared by at least 2 rounds ofultracentrifugation. In some embodiments, the extract is prepared bymore than 2 rounds of untracentrifugation. In some embodiments, theextract is prepared by at least 4 rounds of untracentrifugation. In someembodiments, the composition further comprises apharmaceutically-acceptable diluent, excipient or carrier. In someembodiments, the composition is formulated as a solution, suspension,paste, ointment, oil, emulsion, microemulsion, cream, lotion, gel, orcombination thereof. In some embodiments, the composition is formulatedfor controlled, sustained, or delayed release. In some embodiments, thecomposition is injected into an osteolytic joint. In some embodiments,the composition is administered by a patch. In some embodiments, thepatch is placed on an osteolytic bone or an osteolytic joint. In someembodiments, the composition is administered by an implant. In someembodiments, the implant is placed on an osteolytic bone or anosteolytic joint. In some embodiments, the composition is administeredby an orthopaedic prosthesis. In some embodiments, the composition isadministered by a bone stent. In some embodiments, the composition is asheet and is wrapped around an osteolytic bone or an osteolytic joint.In some embodiments, the methods further comprise administering to theindividual a calcium supplement. In some embodiments, the disease,disorder, or condition characterized by excessive or undesiredosteoclast differentiation is arthritis, osteoporosis, a bone tumor,Paget's Disease, alveolar bone degradation, or any combination thereof.In some embodiments, the arthritis is osteoarthritis, rheumatoidarthritis, psoriatic arthritis, or any combination thereof.

Disclosed herein, in certain embodiments, are methods of inhibiting boneresorption in an individual in need thereof, comprising administering tothe individual a composition comprising substantially isolated HC-HAcomplex. In some embodiments, the HC-HA complex is obtained by a processcomprising: (a) providing a reaction mixture comprising: (i) HA (e.g.,HMW HA); (ii) IαI, wherein the IαI is optionally in serum or isolatedfrom serum; (iii) TSG-6, wherein the TSG-6 is optionally recombinant;and (iv) PTX3, wherein the PTX3 is optionally recombinant; wherein atleast one of HA, IαI, TSG-6, or PTX3 is optionally generated by aplurality of cells present in the reaction mixture; (b) incubating thereaction mixture for a period of time sufficient to produce HC-HAcomplex; and (c) isolating and purifying the HC-HA complex. In someembodiments, the substantially isolated HC-HA complex is derived fromplacenta, umbilical cord, chorion, amnion-chorion, placental amnioticmembrane (PAM), umbilical cord amniotic membrane (UCAM), or anycombinations thereof. In some embodiments, the substantially isolatedHC-HA complex is derived from frozen or previously frozen placentalamniotic membrane (PAM), frozen or previously frozen umbilical cordamniotic membrane (UCAM), frozen or previously frozen placenta, frozenor previously frozen umbilical cord, frozen or previously frozenchorion, frozen or previously frozen amnion-chorion, or any combinationsthereof. In some embodiments, the composition further comprises apharmaceutically-acceptable diluents, excipient or carrier. In someembodiments, the composition is formulated as a solution, suspension,paste, ointment, oil, emulsion, microemulsion, cream, lotion, gel, orcombination thereof. In some embodiments, the composition is formulatedfor controlled, sustained, or delayed release. In some embodiments, thecomposition is injected into an osteolytic joint. In some embodiments,the composition is administered by a patch. In some embodiments, thepatch is placed on an osteolytic bone or an osteolytic joint. In someembodiments, the composition is administered by an implant. In someembodiments, the implant is placed on an osteolytic bone or anosteolytic joint. In some embodiments, the composition is administeredby an orthopaedic prosthesis. In some embodiments, the composition isadministered by a bone stent. In some embodiments, the methods furthercomprise administering to the individual a calcium supplement. In someembodiments, the disease, disorder, or condition characterized byexcessive or undesired osteoclast differentiation is arthritis,osteoporosis, alveolar bone degradation, Paget's Disease, a bone tumor,or any combination thereof.

Disclosed herein, in certain embodiments, are methods of treating adisease, disorder, or condition characterized by excessive or undesiredbone resorption, the method comprising administering to an individual inneed thereof a composition comprising substantially isolated HC-HAcomplex. In some embodiments, the HC-HA complex is obtained by a processcomprising: (a) providing a reaction mixture comprising: (i) HA (e.g.,HMW HA); (ii) IαI, wherein the IαI is optionally in serum or isolatedfrom serum; (iii) TSG-6, wherein the TSG-6 is optionally recombinant;and (iv) PTX3, wherein the PTX3 is optionally recombinant; wherein atleast one of HA, IαI, TSG-6, or PTX3 is optionally generated by aplurality of cells present in the reaction mixture; (b) incubating thereaction mixture for a period of time sufficient to produce HC-HAcomplex; and (c) isolating and purifying the HC-HA complex. In someembodiments, the substantially isolated HC-HA complex is derived fromplacenta, umbilical cord, chorion, amnion-chorion, placental amnioticmembrane (PAM), umbilical cord amniotic membrane (UCAM), or anycombinations thereof. In some embodiments, the substantially isolatedHC-HA complex is derived from frozen or previously frozen placentalamniotic membrane (PAM), frozen or previously frozen umbilical cordamniotic membrane (UCAM), frozen or previously frozen placenta, frozenor previously frozen umbilical cord, frozen or previously frozenchorion, frozen or previously frozen amnion-chorion, or any combinationsthereof. In some embodiments, the composition further comprises apharmaceutically-acceptable diluents, excipient or carrier. In someembodiments, the composition is formulated as a solution, suspension,paste, ointment, oil, emulsion, microemulsion, cream, lotion, gel, orcombination thereof. In some embodiments, the composition is formulatedfor controlled, sustained, or delayed release. In some embodiments, thecomposition is injected into an osteolytic joint. In some embodiments,the composition is administered by a patch. In some embodiments, thepatch is placed on an osteolytic bone or an osteolytic joint. In someembodiments, the composition is administered by an implant. In someembodiments, the implant is placed on an osteolytic bone or anosteolytic joint. In some embodiments, the composition is administeredby an orthopaedic prosthesis. In some embodiments, the composition isadministered by a bone stent. In some embodiments, the methods furthercomprise administering to the individual a calcium supplement. In someembodiments, the disease, disorder, or condition characterized byexcessive or undesired osteoclast differentiation is arthritis,osteoporosis, alveolar bone degradation, Paget's Disease, a bone tumor,or any combination thereof.

Disclosed herein, in certain embodiments, are methods of promoting orinducing bone formation in an individual in need thereof, comprisingadministering to the individual a composition comprising substantiallyisolated HC-HA complex. In some embodiments, the HC-HA complex isobtained by a process comprising: (a) providing a reaction mixturecomprising: (i) HA (e.g., HMW HA); (ii) IαI, wherein the IαI isoptionally in serum or isolated from serum; (iii) TSG-6, wherein theTSG-6 is optionally recombinant; and (iv) PTX3, wherein the PTX3 isoptionally recombinant; wherein at least one of HA, IαI, TSG-6, or PTX3is optionally generated by a plurality of cells present in the reactionmixture; (b) incubating the reaction mixture for a period of timesufficient to produce HC-HA complex; and (c) isolating and purifying theHC-HA complex. In some embodiments, the substantially isolated HC-HAcomplex is derived from placenta, umbilical cord, chorion,amnion-chorion, placental amniotic membrane (PAM), umbilical cordamniotic membrane (UCAM), or any combinations thereof. In someembodiments, the substantially isolated HC-HA complex is derived fromfrozen or previously frozen placental amniotic membrane (PAM), frozen orpreviously frozen umbilical cord amniotic membrane (UCAM), frozen orpreviously frozen placenta, frozen or previously frozen umbilical cord,frozen or previously frozen chorion, frozen or previously frozenamnion-chorion, or any combinations thereof. In some embodiments, thecomposition further comprises a pharmaceutically-acceptable diluents,excipient or carrier. In some embodiments, the composition is formulatedas a solution, suspension, paste, ointment, oil, emulsion,microemulsion, cream, lotion, gel, or combination thereof. In someembodiments, the composition is formulated for controlled, sustained, ordelayed release. In some embodiments, the composition is injected intoan osteolytic joint. In some embodiments, the composition isadministered by a patch. In some embodiments, the patch is placed on anosteolytic bone or an osteolytic joint. In some embodiments, thecomposition is administered by an implant. In some embodiments, theimplant is placed on an osteolytic bone or an osteolytic joint. In someembodiments, the composition is administered by an orthopaedicprosthesis. In some embodiments, the composition is administered by abone stent. In some embodiments, the methods further compriseadministering to the individual a calcium supplement. In someembodiments, the disease, disorder, or condition characterized byexcessive or undesired osteoclast differentiation is arthritis,osteoporosis, alveolar bone degradation, Paget's Disease, a bone tumor,or any combination thereof.

Disclosed herein, in certain embodiments, are methods of treating adisease, disorder or condition characterized by a deficiency of boneformation in an individual in need thereof, the method comprisingadministering to an individual in need thereof a composition comprisingsubstantially isolated HC-HA complex. In some embodiments, the HC-HAcomplex is obtained by a process comprising: (a) providing a reactionmixture comprising: ((i) HA (e.g., HMW HA); (ii) IαI, wherein the IαI isoptionally in serum or isolated from serum; (iii) TSG-6, wherein theTSG-6 is optionally recombinant; and (iv) PTX3, wherein the PTX3 isoptionally recombinant; wherein at least one of HA, IαI, TSG-6, or PTX3is optionally generated by a plurality of cells present in the reactionmixture; (b) incubating the reaction mixture for a period of timesufficient to produce HC-HA complex; and (c) isolating and purifying theHC-HA complex. In some embodiments, the substantially isolated HC-HAcomplex is derived from placenta, umbilical cord, chorion,amnion-chorion, placental amniotic membrane (PAM), umbilical cordamniotic membrane (UCAM), or any combinations thereof. In someembodiments, the substantially isolated HC-HA complex is derived fromfrozen or previously frozen placental amniotic membrane (PAM), frozen orpreviously frozen umbilical cord amniotic membrane (UCAM), frozen orpreviously frozen placenta, frozen or previously frozen umbilical cord,frozen or previously frozen chorion, frozen or previously frozenamnion-chorion, or any combinations thereof. In some embodiments, thecomposition further comprises a pharmaceutically-acceptable diluents,excipient or carrier. In some embodiments, the composition is formulatedas a solution, suspension, paste, ointment, oil, emulsion,microemulsion, cream, lotion, gel, or combination thereof. In someembodiments, the composition is formulated for controlled, sustained, ordelayed release. In some embodiments, the composition is injected intoan osteolytic joint. In some embodiments, the composition isadministered by a patch. In some embodiments, the patch is placed on anosteolytic bone or an osteolytic joint. In some embodiments, thecomposition is administered by an implant. In some embodiments, theimplant is placed on an osteolytic bone or an osteolytic joint. In someembodiments, the composition is administered by an orthopaedicprosthesis. In some embodiments, the composition is administered by abone stent. In some embodiments, the methods further compriseadministering to the individual a calcium supplement. In someembodiments, the disease, disorder, or condition characterized byexcessive or undesired osteoclast differentiation is arthritis,osteoporosis, alveolar bone degradation, Paget's Disease, a bone tumor,or any combination thereof.

Disclosed herein, in certain embodiments, are patches comprising fetalsupport tissue product or an extract thereof. In some embodiments, thefetal support tissue product comprises placental amniotic membrane(PAM), umbilical cord amniotic membrane (UCAM), placenta, umbilicalcord, chorion, amnion-chorion, or any combinations thereof. In someembodiments, the fetal support tissue product comprises frozen orpreviously frozen placental amniotic membrane (PAM), frozen orpreviously frozen umbilical cord amniotic membrane (UCAM), frozen orpreviously frozen placenta, frozen or previously frozen umbilical cord,frozen or previously frozen chorion, frozen or previously frozenamnion-chorion, or any combinations thereof. In some embodiments, thefetal support tissue product in the form of a pulverized powder or ahomogenate. In some embodiments, the extract comprises HC-HA.

Disclosed herein, in certain embodiments, are patches comprisingsubstantially isolated HC-HA. In some embodiments, the substantiallyisolated HC-HA complex is provided in a composition comprising fetalsupport tissue product or in an extract of fetal support tissue product.In some embodiments, the substantially isolated HC-HA complex isobtained by a process comprising: (a) providing a reaction mixturecomprising: (i) HA; (ii) IαI, wherein the IαI is optionally in serum orisolated from serum; (iii) TSG-6, wherein the TSG-6 is optionallyrecombinant; and (iv) PTX3, wherein the PTX3 is optionally recombinant;wherein at least one of HA, IαI, TSG-6, or PTX3 is optionally generatedby a plurality of cells present in the reaction mixture; (b) incubatingthe reaction mixture for a period of time sufficient to produce HC-HAcomplex; and (c) isolating and purifying the HC-HA complex. In someembodiments, the substantially isolated HC-HA complex is derived fromplacenta, umbilical cord, chorion, amnion-chorion, placental amnioticmembrane (PAM), umbilical cord amniotic membrane (UCAM), or anycombinations thereof. In some embodiments, the substantially isolatedHC-HA complex is derived from frozen or previously frozen placentalamniotic membrane (PAM), frozen or previously frozen umbilical cordamniotic membrane (UCAM), frozen or previously frozen placenta, frozenor previously frozen umbilical cord, frozen or previously frozenchorion, frozen or previously frozen amnion-chorion, or any combinationsthereof.

Disclosed herein, in certain embodiments, are bone implants comprisingfetal support tissue product or an extract thereof. In some embodiments,the bone implant is a bone stent or an osteointegrated implant. In someembodiments, the fetal support tissue product or the extract thereof iscoated onto the outside of the bone implant. In some embodiments, thefetal support tissue product or the extract thereof elutes from the boneimplant. In some embodiments, the fetal support tissue product comprisesplacental amniotic membrane (PAM), umbilical cord amniotic membrane(UCAM), placenta, umbilical cord, chorion, amnion-chorion, or anycombinations thereof. In some embodiments, the fetal support tissueproduct comprises frozen or previously frozen placental amnioticmembrane (PAM), frozen or previously frozen umbilical cord amnioticmembrane (UCAM), frozen or previously frozen placenta, frozen orpreviously frozen umbilical cord, frozen or previously frozen chorion,frozen or previously frozen amnion-chorion, or any combinations thereof.In some embodiments, the fetal support tissue product in the form of apulverized powder or a homogenate. In some embodiments, the extractcomprises HC-HA.

Disclosed herein, in certain embodiments, are bone implants comprisingsubstantially isolated HC-HA. In some embodiments, the bone implant is abone stent or an osteointegrated implant. In some embodiments, thesubstantially isolated HC-HA is coated onto the outside of the boneimplant. In some embodiments, the substantially isolated HC-HA elutesfrom the bone implant. In some embodiments, the substantially isolatedHC-HA complex is provided in a composition comprising fetal supporttissue product or in an extract of fetal support tissue product. In someembodiments, the substantially isolated HC-HA complex is obtained by aprocess comprising: (a) providing a reaction mixture comprising: (i) HA;(ii) IαI, wherein the IαI is optionally in serum or isolated from serum;(iii) TSG-6, wherein the TSG-6 is optionally recombinant; and (iv) PTX3,wherein the PTX3 is optionally recombinant; wherein at least one of HA,IαI, TSG-6, or PTX3 is optionally generated by a plurality of cellspresent in the reaction mixture; (b) incubating the reaction mixture fora period of time sufficient to produce HC-HA complex; and (c) isolatingand purifying the HC-HA complex. In some embodiments, the substantiallyisolated HC-HA complex is derived from placenta, umbilical cord,chorion, amnion-chorion, placental amniotic membrane (PAM), umbilicalcord amniotic membrane (UCAM), or any combinations thereof. In someembodiments, the substantially isolated HC-HA complex is derived fromfrozen or previously frozen placental amniotic membrane (PAM), frozen orpreviously frozen umbilical cord amniotic membrane (UCAM), frozen orpreviously frozen placenta, frozen or previously frozen umbilical cord,frozen or previously frozen chorion, frozen or previously frozenamnion-chorion, or any combinations thereof.

Disclosed herein, in certain embodiments, are compositions foradministration to a bone comprising fetal support tissue product or anextract thereof. In some embodiments, the composition is administered byinjection, a bone implant or a patch. In some embodiments, the boneimplant is a bone stent. In some embodiments, the fetal support tissueproduct or the extract thereof is coated onto the outside of a boneimplant. In some embodiments, the fetal support tissue product or theextract thereof elutes from the bone implant. In some embodiments, thefetal support tissue product comprises placental amniotic membrane(PAM), umbilical cord amniotic membrane (UCAM), placenta, umbilicalcord, chorion, amnion-chorion, or any combinations thereof. In someembodiments, the fetal support tissue product comprises frozen orpreviously frozen placental amniotic membrane (PAM), frozen orpreviously frozen umbilical cord amniotic membrane (UCAM), frozen orpreviously frozen placenta, frozen or previously frozen umbilical cord,frozen or previously frozen chorion, frozen or previously frozenamnion-chorion, or any combinations thereof. In some embodiments, thefetal support tissue product in the form of a pulverized powder or ahomogenate. In some embodiments, the extract comprises HC-HA.

Disclosed herein, in certain embodiments, are compositions foradministration to a bone comprising substantially isolated HC-HA. Insome embodiments, the composition is administered by injection, a boneimplant or a patch. In some embodiments, the bone implant is a bonestent. In some embodiments, the fetal support tissue product or theextract thereof is coated onto the outside of a bone implant. In someembodiments, the fetal support tissue product or the extract thereofelutes from the bone implant. In some embodiments, the substantiallyisolated HC-HA complex is provided in a composition comprising fetalsupport tissue product or in an extract of fetal support tissue product.In some embodiments, the substantially isolated HC-HA complex isobtained by a process comprising: (a) providing a reaction mixturecomprising: (i) HA; (ii) IαI, wherein the IαI is optionally in serum orisolated from serum; (iii) TSG-6, wherein the TSG-6 is optionallyrecombinant; and (iv) PTX3, wherein the PTX3 is optionally recombinant;wherein at least one of HA, IαI, TSG-6, or PTX3 is optionally generatedby a plurality of cells present in the reaction mixture; (b) incubatingthe reaction mixture for a period of time sufficient to produce HC-HAcomplex; and (c) isolating and purifying the HC-HA complex. In someembodiments, the substantially isolated HC-HA complex is derived fromplacenta, umbilical cord, chorion, amnion-chorion, placental amnioticmembrane (PAM), umbilical cord amniotic membrane (UCAM), or anycombinations thereof. In some embodiments, the substantially isolatedHC-HA complex is derived from frozen or previously frozen placentalamniotic membrane (PAM), frozen or previously frozen umbilical cordamniotic membrane (UCAM), frozen or previously frozen placenta, frozenor previously frozen umbilical cord, frozen or previously frozenchorion, frozen or previously frozen amnion-chorion, or any combinationsthereof.

Disclosed herein, in certain embodiments, are methods of inhibitingosteoclast differentiation in an individual in need thereof, comprisingadministering to the individual a composition comprising product fetalsupport tissue product or an extract thereof. In some embodiments, thefetal support tissue product is substantially isolated placentalamniotic membrane (PAM), umbilical cord amniotic membrane (UCAM),placenta, umbilical cord, chorion, amnion-chorion, or any combinationsthereof. In some embodiments, the fetal support tissue product is frozenor previously frozen PAM, frozen or previously frozen UCAM, frozen orpreviously frozen placenta, frozen or previously frozen umbilical cord,frozen or previously frozen chorion, frozen or previously frozenamnion-chorion, or any combinations thereof. In some embodiments, thefetal support tissue product is cryopreserved, lyophilized, terminallysterilized, or a combination thereof. In some embodiments, the fetalsupport tissue product is in the form of a pulverized powder or ahomogenate. In some embodiments, the fetal support tissue product doesnot comprise a vein or an artery, a cell with metabolic activity, activeHIV-1, active HIV-2, active HTLV-1, active hepatitis B, active hepatitisC, active West Nile Virus, active cytomegalovirus, active humantransmissible spongiform encephalopathy, or active treponema pallidum.In some embodiments, the fetal support tissue product is obtained from ahuman, non-primate human, cow or pig. In some embodiments, thecomposition further comprises a pharmaceutically-acceptable diluents,excipient or carrier. In some embodiments, the composition is formulatedas a solution, suspension, paste, ointment, oil, emulsion,microemulsion, cream, lotion, gel, or combination thereof. In someembodiments, the composition comprises substantially-isolated tissueproduct formulated into microspheres, microparticles, or liposomes. Insome embodiments, the composition is formulated into PLGA copolymers. Insome embodiments, the composition is formulated for controlled,sustained, or delayed release. In some embodiments, the composition isinjected into an osteolytic joint. In some embodiments, the compositionis administered by a patch. In some embodiments, the patch is placed onan osteolytic bone or an osteolytic joint. In some embodiments, thecomposition is administered by an implant. In some embodiments, thecomposition is formulated into an orthopaedic prosthesis. In someembodiments, the fetal support tissue product is in the form of a sheet.In some embodiments, the composition further comprises a backing. Insome embodiments, the composition is wrapped around an osteolytic boneor an osteolytic joint. In some embodiments, the methods furthercomprise administering to the individual a calcium supplement.

Disclosed herein, in certain embodiments, are methods of promotingmineralization by osteoblasts in an individual in need thereof,comprising administering to the individual a composition comprising afetal support tissue product or an extract thereof. In some embodiments,the fetal support tissue product is substantially isolated placentalamniotic membrane (PAM), umbilical cord amniotic membrane (UCAM),placenta, umbilical cord, chorion, amnion-chorion, or any combinationsthereof. In some embodiments, the fetal support tissue product is frozenor previously frozen PAM, frozen or previously frozen UCAM, frozen orpreviously frozen placenta, frozen or previously frozen umbilical cord,frozen or previously frozen chorion, frozen or previously frozenamnion-chorion, or any combinations thereof. In some embodiments, thefetal support tissue product is cryopreserved, lyophilized, terminallysterilized, or a combination thereof. In some embodiments, the fetalsupport tissue product is in the form of a pulverized powder or ahomogenate. In some embodiments, the fetal support tissue product doesnot comprise a vein or an artery, a cell with metabolic activity, activeHIV-1, active HIV-2, active HTLV-1, active hepatitis B, active hepatitisC, active West Nile Virus, active cytomegalovirus, active humantransmissible spongiform encephalopathy, or active treponema pallidum.In some embodiments, the fetal support tissue product is obtained from ahuman, non-primate human, cow or pig. In some embodiments, thecomposition further comprises a pharmaceutically-acceptable diluents,excipient or carrier. In some embodiments, the composition is formulatedas a solution, suspension, paste, ointment, oil, emulsion,microemulsion, cream, lotion, gel, or combination thereof. In someembodiments, the composition comprises substantially-isolated tissueproduct formulated into microspheres, microparticles, or liposomes. Insome embodiments, the composition is formulated into PLGA copolymers. Insome embodiments, the composition is formulated for controlled,sustained, or delayed release. In some embodiments, the composition isinjected into an osteolytic joint. In some embodiments, the compositionis administered by a patch. In some embodiments, the patch is placed onan osteolytic bone or an osteolytic joint. In some embodiments, thecomposition is administered by an implant. In some embodiments, thecomposition is formulated into an orthopaedic prosthesis. In someembodiments, the fetal support tissue product is in the form of a sheet.In some embodiments, the composition further comprises a backing. Insome embodiments, the composition is wrapped around an osteolytic boneor an osteolytic joint. In some embodiments, the methods furthercomprise administering to the individual a calcium supplement.

Disclosed herein, in certain embodiments, are methods of inhibiting boneresorption in an individual in need thereof, comprising administering tothe individual a composition comprising a fetal support tissue productor an extract thereof. In some embodiments, the fetal support tissueproduct is substantially isolated placental amniotic membrane (PAM),umbilical cord amniotic membrane (UCAM), placenta, umbilical cord,chorion, amnion-chorion, or any combinations thereof. In someembodiments, the fetal support tissue product is frozen or previouslyfrozen PAM, frozen or previously frozen UCAM, frozen or previouslyfrozen placenta, frozen or previously frozen umbilical cord, frozen orpreviously frozen chorion, frozen or previously frozen amnion-chorion,or any combinations thereof. In some embodiments, the fetal supporttissue product is cryopreserved, lyophilized, terminally sterilized, ora combination thereof. In some embodiments, the fetal support tissueproduct is in the form of a pulverized powder or a homogenate. In someembodiments, the fetal support tissue product does not comprise a veinor an artery, a cell with metabolic activity, active HIV-1, activeHIV-2, active HTLV-1, active hepatitis B, active hepatitis C, activeWest Nile Virus, active cytomegalovirus, active human transmissiblespongiform encephalopathy, or active treponema pallidum. In someembodiments, the fetal support tissue product is obtained from a human,non-primate human, cow or pig. In some embodiments, the compositionfurther comprises a pharmaceutically-acceptable diluents, excipient orcarrier. In some embodiments, the composition is formulated as asolution, suspension, paste, ointment, oil, emulsion, microemulsion,cream, lotion, gel, or combination thereof. In some embodiments, thecomposition comprises substantially-isolated tissue product formulatedinto microspheres, microparticles, or liposomes. In some embodiments,the composition is formulated into PLGA copolymers. In some embodiments,the composition is formulated for controlled, sustained, or delayedrelease. In some embodiments, the composition is injected into anosteolytic joint. In some embodiments, the composition is administeredby a patch. In some embodiments, the patch is placed on an osteolyticbone or an osteolytic joint. In some embodiments, the composition isadministered by an implant. In some embodiments, the composition isformulated into an orthopaedic prosthesis. In some embodiments, thefetal support tissue product is in the form of a substantially-flattenedsheet. In some embodiments, the composition further comprises a backing.In some embodiments, the composition is wrapped around an osteolyticbone or an osteolytic joint. In some embodiments, the methods furthercomprise administering to the individual a calcium supplement.

Disclosed herein, in certain embodiments, are methods of inhibiting boneremodeling in an individual in need thereof, comprising administering tothe individual a composition comprising a fetal support tissue productor an extract thereof. In some embodiments, the fetal support tissueproduct is substantially isolated placental amniotic membrane (PAM),umbilical cord amniotic membrane (UCAM), placenta, umbilical cord,chorion, amnion-chorion, or any combinations thereof. In someembodiments, the fetal support tissue product is frozen or previouslyfrozen PAM, frozen or previously frozen UCAM, frozen or previouslyfrozen placenta, frozen or previously frozen umbilical cord, frozen orpreviously frozen chorion, frozen or previously frozen amnion-chorion,or any combinations thereof. In some embodiments, the fetal supporttissue product is cryopreserved, lyophilized, terminally sterilized, ora combination thereof. In some embodiments, the fetal support tissueproduct is in the form of a pulverized powder or a homogenate. In someembodiments, the fetal support tissue product does not comprise a veinor an artery, a cell with metabolic activity, active HIV-1, activeHIV-2, active HTLV-1, active hepatitis B, active hepatitis C, activeWest Nile Virus, active cytomegalovirus, active human transmissiblespongiform encephalopathy, or active treponema pallidum. In someembodiments, the fetal support tissue product is obtained from a human,non-primate human, cow or pig. In some embodiments, the compositionfurther comprises a pharmaceutically-acceptable diluents, excipient orcarrier. In some embodiments, the composition is formulated as asolution, suspension, paste, ointment, oil, emulsion, microemulsion,cream, lotion, gel, or combination thereof. In some embodiments, thecomposition comprises substantially-isolated tissue product formulatedinto microspheres, microparticles, or liposomes. In some embodiments,the composition is formulated into PLGA copolymers. In some embodiments,the composition is formulated for controlled, sustained, or delayedrelease. In some embodiments, the composition is injected into anosteolytic joint. In some embodiments, the composition is administeredby a patch. In some embodiments, the patch is placed on an osteolyticbone or an osteolytic joint. In some embodiments, the composition isadministered by an implant. In some embodiments, the composition isformulated into an orthopaedic prosthesis. In some embodiments, thefetal support tissue product is in the form of a substantially-flattenedsheet. In some embodiments, the composition further comprises a backing.In some embodiments, the composition is wrapped around an osteolyticbone or an osteolytic joint. In some embodiments, the methods furthercomprise administering to the individual a calcium supplement.

Disclosed herein, in certain embodiments, are methods of balancing boneresorption and bone formation, comprising administering to an individualin need thereof a therapeutically-effective amount of a compositioncomprising a fetal support tissue product or an extract thereof. In someembodiments, the fetal support tissue product is substantially isolatedplacental amniotic membrane (PAM), umbilical cord amniotic membrane(UCAM), placenta, umbilical cord, chorion, amnion-chorion, or anycombinations thereof. In some embodiments, the fetal support tissueproduct is frozen or previously frozen PAM, frozen or previously frozenUCAM, frozen or previously frozen placenta, frozen or previously frozenumbilical cord, frozen or previously frozen chorion, frozen orpreviously frozen amnion-chorion, or any combinations thereof. In someembodiments, the fetal support tissue product is cryopreserved,lyophilized, terminally sterilized, or a combination thereof. In someembodiments, the fetal support tissue product is in the form of apulverized powder or a homogenate. In some embodiments, the fetalsupport tissue product does not comprise a vein or an artery, a cellwith metabolic activity, active HIV-1, active HIV-2, active HTLV-1,active hepatitis B, active hepatitis C, active West Nile Virus, activecytomegalovirus, active human transmissible spongiform encephalopathy,or active treponema pallidum. In some embodiments, the fetal supporttissue product is obtained from a human, non-primate human, cow or pig.In some embodiments, the composition further comprises apharmaceutically-acceptable diluents, excipient or carrier. In someembodiments, the composition is formulated as a solution, suspension,paste, ointment, oil, emulsion, microemulsion, cream, lotion, gel, orcombination thereof. In some embodiments, the composition comprisessubstantially-isolated tissue product formulated into microspheres,microparticles, or liposomes. In some embodiments, the composition isformulated into PLGA copolymers. In some embodiments, the composition isformulated for controlled, sustained, or delayed release. In someembodiments, the composition is injected into an osteolytic joint. Insome embodiments, the composition is administered by a patch. In someembodiments, the patch is placed on an osteolytic bone or an osteolyticjoint. In some embodiments, the composition is administered by animplant. In some embodiments, the composition is formulated into anorthopaedic prosthesis. In some embodiments, the fetal support tissueproduct is in the form of a substantially-flattened sheet. In someembodiments, the composition further comprises a backing. In someembodiments, the composition is wrapped around an osteolytic bone or anosteolytic joint. In some embodiments, the methods further compriseadministering to the individual a calcium supplement.

Disclosed herein, in certain embodiments, are methods of treating adisease, disorder, or condition characterized by excessive or undesiredosteoclast differentiation, the methods comprising administering to anindividual in need thereof a therapeutically-effective amount of acomposition comprising a fetal support tissue product or an extractthereof. In some embodiments, the fetal support tissue product issubstantially isolated placental amniotic membrane (PAM), umbilical cordamniotic membrane (UCAM), placenta, umbilical cord, chorion,amnion-chorion, or any combinations thereof. In some embodiments, thefetal support tissue product is frozen or previously frozen PAM, frozenor previously frozen UCAM, frozen or previously frozen placenta, frozenor previously frozen umbilical cord, frozen or previously frozenchorion, frozen or previously frozen amnion-chorion, or any combinationsthereof. In some embodiments, the fetal support tissue product iscryopreserved, lyophilized, terminally sterilized, or a combinationthereof. In some embodiments, the fetal support tissue product is in theform of a pulverized powder or a homogenate. In some embodiments, thefetal support tissue product does not comprise a vein or an artery, acell with metabolic activity, active HIV-1, active HIV-2, active HTLV-1,active hepatitis B, active hepatitis C, active West Nile Virus, activecytomegalovirus, active human transmissible spongiform encephalopathy,or active treponema pallidum. In some embodiments, the fetal supporttissue product is obtained from a human, non-primate human, cow or pig.In some embodiments, the composition further comprises apharmaceutically-acceptable diluents, excipient or carrier. In someembodiments, the composition is formulated as a solution, suspension,paste, ointment, oil, emulsion, microemulsion, cream, lotion, gel, orcombination thereof. In some embodiments, the composition comprisessubstantially-isolated tissue product formulated into microspheres,microparticles, or liposomes. In some embodiments, the composition isformulated into PLGA copolymers. In some embodiments, the composition isformulated for controlled, sustained, or delayed release. In someembodiments, the composition is injected into an osteolytic joint. Insome embodiments, the composition is administered by a patch. In someembodiments, the patch is placed on an osteolytic bone or an osteolyticjoint. In some embodiments, the composition is administered by animplant. In some embodiments, the composition is formulated into anorthopaedic prosthesis. In some embodiments, the fetal support tissueproduct is in the form of a substantially-flattened sheet. In someembodiments, the composition further comprises a backing. In someembodiments, the composition is wrapped around an osteolytic bone or anosteolytic joint. In some embodiments, the methods further compriseadministering to the individual a calcium supplement.

Disclosed herein, in certain embodiments, are methods of treating adisease, disorder, or condition characterized by excessive or undesiredbone absorption by osteoclasts, the methods comprising administering toan individual in need thereof a therapeutically-effective amount of acomposition comprising a fetal support tissue product or an extractthereof. In some embodiments, the fetal support tissue product issubstantially isolated placental amniotic membrane (PAM), umbilical cordamniotic membrane (UCAM), placenta, umbilical cord, chorion,amnion-chorion, or any combinations thereof. In some embodiments, thefetal support tissue product is frozen or previously frozen PAM, frozenor previously frozen UCAM, frozen or previously frozen placenta, frozenor previously frozen umbilical cord, frozen or previously frozenchorion, frozen or previously frozen amnion-chorion, or any combinationsthereof. In some embodiments, the fetal support tissue product iscryopreserved, lyophilized, terminally sterilized, or a combinationthereof. In some embodiments, the fetal support tissue product is in theform of a pulverized powder or a homogenate. In some embodiments, thefetal support tissue product does not comprise a vein or an artery, acell with metabolic activity, active HIV-1, active HIV-2, active HTLV-1,active hepatitis B, active hepatitis C, active West Nile Virus, activecytomegalovirus, active human transmissible spongiform encephalopathy,or active treponema pallidum. In some embodiments, the fetal supporttissue product is obtained from a human, non-primate human, cow or pig.In some embodiments, the composition further comprises apharmaceutically-acceptable diluents, excipient or carrier. In someembodiments, the composition is formulated as a solution, suspension,paste, ointment, oil, emulsion, microemulsion, cream, lotion, gel, orcombination thereof. In some embodiments, the composition comprisessubstantially-isolated tissue product formulated into microspheres,microparticles, or liposomes. In some embodiments, the composition isformulated into PLGA copolymers. In some embodiments, the composition isformulated for controlled, sustained, or delayed release. In someembodiments, the composition is injected into an osteolytic joint. Insome embodiments, the composition is administered by a patch. In someembodiments, the patch is placed on an osteolytic bone or an osteolyticjoint. In some embodiments, the composition is administered by animplant. In some embodiments, the composition is formulated into anorthopaedic prosthesis. In some embodiments, the fetal support tissueproduct is in the form of a substantially-flattened sheet. In someembodiments, the composition further comprises a backing. In someembodiments, the composition is wrapped around an osteolytic bone or anosteolytic joint. In some embodiments, the methods further compriseadministering to the individual a calcium supplement.

Disclosed herein, in certain embodiments, are methods of treating adisease, disorder, or condition characterized by deficient or defectivebone formation, the methods comprising administering to an individual inneed thereof a therapeutically-effective amount of a compositioncomprising a fetal support tissue product or an extract thereof. In someembodiments, the fetal support tissue product is substantially isolatedplacental amniotic membrane (PAM), umbilical cord amniotic membrane(UCAM), placenta, umbilical cord, chorion, amnion-chorion, or anycombinations thereof. In some embodiments, the fetal support tissueproduct is frozen or previously frozen PAM, frozen or previously frozenUCAM, frozen or previously frozen placenta, frozen or previously frozenumbilical cord, frozen or previously frozen chorion, frozen orpreviously frozen amnion-chorion, or any combinations thereof. In someembodiments, the fetal support tissue product is cryopreserved,lyophilized, terminally sterilized, or a combination thereof. In someembodiments, the fetal support tissue product is in the form of apulverized powder or a homogenate. In some embodiments, the fetalsupport tissue product does not comprise a vein or an artery, a cellwith metabolic activity, active HIV-1, active HIV-2, active HTLV-1,active hepatitis B, active hepatitis C, active West Nile Virus, activecytomegalovirus, active human transmissible spongiform encephalopathy,or active treponema pallidum. In some embodiments, the fetal supporttissue product is obtained from a human, non-primate human, cow or pig.In some embodiments, the composition further comprises apharmaceutically-acceptable diluents, excipient or carrier. In someembodiments, the composition is formulated as a solution, suspension,paste, ointment, oil, emulsion, microemulsion, cream, lotion, gel, orcombination thereof. In some embodiments, the composition comprisessubstantially-isolated tissue product formulated into microspheres,microparticles, or liposomes. In some embodiments, the composition isformulated into PLGA copolymers. In some embodiments, the composition isformulated for controlled, sustained, or delayed release. In someembodiments, the composition is injected into an osteolytic joint. Insome embodiments, the composition is administered by a patch. In someembodiments, the patch is placed on an osteolytic bone or an osteolyticjoint. In some embodiments, the composition is administered by animplant. In some embodiments, the composition is formulated into anorthopaedic prosthesis. In some embodiments, the fetal support tissueproduct is in the form of a substantially-flattened sheet. In someembodiments, the composition further comprises a backing. In someembodiments, the composition is wrapped around an osteolytic bone or anosteolytic joint. In some embodiments, the methods further compriseadministering to the individual a calcium supplement.

Disclosed herein, in certain embodiments, are methods of treatingarthritis, the methods comprising administering to an individual in needthereof a therapeutically-effective amount of a composition comprising afetal support tissue product or an extract thereof. In some embodiments,the arthritis is osteoarthritis, rheumatoid arthritis, psoriaticarthritis, or any combination thereof. In some embodiments, the fetalsupport tissue product is substantially isolated placental amnioticmembrane (PAM), umbilical cord amniotic membrane (UCAM), placenta,umbilical cord, chorion, amnion-chorion, or any combinations thereof. Insome embodiments, the fetal support tissue product is frozen orpreviously frozen PAM, frozen or previously frozen UCAM, frozen orpreviously frozen placenta, frozen or previously frozen umbilical cord,frozen or previously frozen chorion, frozen or previously frozenamnion-chorion, or any combinations thereof. In some embodiments, thefetal support tissue product is cryopreserved, lyophilized, terminallysterilized, or a combination thereof. In some embodiments, the fetalsupport tissue product is in the form of a pulverized powder or ahomogenate. In some embodiments, the fetal support tissue product doesnot comprise a vein or an artery, a cell with metabolic activity, activeHIV-1, active HIV-2, active HTLV-1, active hepatitis B, active hepatitisC, active West Nile Virus, active cytomegalovirus, active humantransmissible spongiform encephalopathy, or active treponema pallidum.In some embodiments, the fetal support tissue product is obtained from ahuman, non-primate human, cow or pig. In some embodiments, thecomposition further comprises a pharmaceutically-acceptable diluents,excipient or carrier. In some embodiments, the composition is formulatedas a solution, suspension, paste, ointment, oil, emulsion,microemulsion, cream, lotion, gel, or combination thereof. In someembodiments, the composition comprises substantially-isolated tissueproduct formulated into microspheres, microparticles, or liposomes. Insome embodiments, the composition is formulated into PLGA copolymers. Insome embodiments, the composition is formulated for controlled,sustained, or delayed release. In some embodiments, the composition isinjected into an osteolytic joint. In some embodiments, the compositionis administered by a patch. In some embodiments, the patch is placed onan osteolytic bone or an osteolytic joint. In some embodiments, thecomposition is administered by an implant. In some embodiments, theimplant is placed on an osteolytic bone or an osteolytic joint. In someembodiments, the composition is formulated into an orthopaedicprosthesis. In some embodiments, the fetal support tissue product is inthe form of a substantially-flattened sheet. In some embodiments, thecomposition further comprises a backing. In some embodiments, thecomposition is wrapped around an osteolytic bone or an osteolytic joint.In some embodiments, the methods further comprise administering to theindividual a calcium supplement. In some embodiments, the methodsfurther comprise administering to the individual an NSAID, acorticosteroid, hyaluronan injections, a DMARD, an analgesic, or anycombination thereof.

Disclosed herein, in certain embodiments, are methods of treatingosteoporosis, the methods comprising administering to an individual inneed thereof a therapeutically-effective amount of a compositioncomprising a fetal support tissue product or an extract thereof. In someembodiments, the fetal support tissue product is substantially isolatedplacental amniotic membrane (PAM), umbilical cord amniotic membrane(UCAM), placenta, umbilical cord, chorion, amnion-chorion, or anycombinations thereof. In some embodiments, the fetal support tissueproduct is frozen or previously frozen PAM, frozen or previously frozenUCAM, frozen or previously frozen placenta, frozen or previously frozenumbilical cord, frozen or previously frozen chorion, frozen orpreviously frozen amnion-chorion, or any combinations thereof. In someembodiments, the fetal support tissue product is cryopreserved,lyophilized, terminally sterilized, or a combination thereof. In someembodiments, the fetal support tissue product is in the form of apulverized powder or a homogenate. In some embodiments, the fetalsupport tissue product does not comprise a vein or an artery, a cellwith metabolic activity, active HIV-1, active HIV-2, active HTLV-1,active hepatitis B, active hepatitis C, active West Nile Virus, activecytomegalovirus, active human transmissible spongiform encephalopathy,or active treponema pallidum. In some embodiments, the fetal supporttissue product is obtained from a human, non-primate human, cow or pig.In some embodiments, the composition further comprises apharmaceutically-acceptable diluents, excipient or carrier. In someembodiments, the composition is formulated as a solution, suspension,paste, ointment, oil, emulsion, microemulsion, cream, lotion, gel, orcombination thereof. In some embodiments, the composition comprisessubstantially-isolated tissue product formulated into microspheres,microparticles, or liposomes. In some embodiments, the composition isformulated into PLGA copolymers. In some embodiments, the composition isformulated for controlled, sustained, or delayed release. In someembodiments, the composition is injected into an osteolytic joint. Insome embodiments, the composition is administered by a patch. In someembodiments, the patch is placed on an osteolytic bone or an osteolyticjoint. In some embodiments, the composition is administered by animplant. In some embodiments, the implant is placed on an osteolyticbone or an osteolytic joint. In some embodiments, the composition isformulated into an orthopaedic prosthesis. In some embodiments, thefetal support tissue product is in the form of a substantially-flattenedsheet. In some embodiments, the composition further comprises a backing.In some embodiments, the composition is wrapped around an osteolyticbone or an osteolytic joint. In some embodiments, the methods furthercomprise administering to the individual a calcium supplement. In someembodiments, the methods further comprise administering to theindividual a bisphosphonate, an estrogen analog, Raloxifene, Calcitonin,Teriparatide, calcium salts, sodium fluoride, RANKL inhibitors,Strontium ranelate, or any combination thereof.

Disclosed herein, in certain embodiments, are methods of treatingalveolar bone degradation, the methods comprising administering to anindividual in need thereof a therapeutically-effective amount of acomposition comprising a fetal support tissue product or an extractthereof. In some embodiments, the fetal support tissue product issubstantially isolated placental amniotic membrane (PAM), umbilical cordamniotic membrane (UCAM), placenta, umbilical cord, chorion,amnion-chorion, or any combinations thereof. In some embodiments, thefetal support tissue product is frozen or previously frozen PAM, frozenor previously frozen UCAM, frozen or previously frozen placenta, frozenor previously frozen umbilical cord, frozen or previously frozenchorion, frozen or previously frozen amnion-chorion, or any combinationsthereof. In some embodiments, the fetal support tissue product iscryopreserved, lyophilized, terminally sterilized, or a combinationthereof. In some embodiments, the fetal support tissue product is in theform of a pulverized powder or a homogenate. In some embodiments, thefetal support tissue product does not comprise a vein or an artery, acell with metabolic activity, active HIV-1, active HIV-2, active HTLV-1,active hepatitis B, active hepatitis C, active West Nile Virus, activecytomegalovirus, active human transmissible spongiform encephalopathy,or active treponema pallidum. In some embodiments, the fetal supporttissue product is obtained from a human, non-primate human, cow or pig.In some embodiments, the composition further comprises apharmaceutically-acceptable diluents, excipient or carrier. In someembodiments, the composition is formulated as a solution, suspension,paste, ointment, oil, emulsion, microemulsion, cream, lotion, gel, orcombination thereof. In some embodiments, the composition comprisessubstantially-isolated tissue product formulated into microspheres,microparticles, or liposomes. In some embodiments, the composition isformulated into PLGA copolymers. In some embodiments, the composition isformulated for controlled, sustained, or delayed release. In someembodiments, the composition is administered by a patch. In someembodiments, the patch is placed on the alveolar bone. In someembodiments, the composition is administered by an implant. In someembodiments, the implant is placed on the alveolar bone. In someembodiments, the fetal support tissue product is in the form of asubstantially-flattened sheet. In some embodiments, the compositionfurther comprises a backing. In some embodiments, the composition isplaced on the alveolar bone.

Disclosed herein, in certain embodiments, are methods of treatingPaget's disease, the methods comprising administering to an individualin need thereof a therapeutically-effective amount of a compositioncomprising a fetal support tissue product or an extract thereof. In someembodiments, the fetal support tissue product is substantially isolatedplacental amniotic membrane (PAM), umbilical cord amniotic membrane(UCAM), placenta, umbilical cord, chorion, amnion-chorion, or anycombinations thereof. In some embodiments, the fetal support tissueproduct is frozen or previously frozen PAM, frozen or previously frozenUCAM, frozen or previously frozen placenta, frozen or previously frozenumbilical cord, frozen or previously frozen chorion, frozen orpreviously frozen amnion-chorion, or any combinations thereof. In someembodiments, the fetal support tissue product is cryopreserved,lyophilized, terminally sterilized, or a combination thereof. In someembodiments, the fetal support tissue product is in the form of apulverized powder or a homogenate. In some embodiments, the fetalsupport tissue product does not comprise a vein or an artery, a cellwith metabolic activity, active HIV-1, active HIV-2, active HTLV-1,active hepatitis B, active hepatitis C, active West Nile Virus, activecytomegalovirus, active human transmissible spongiform encephalopathy,or active treponema pallidum. In some embodiments, the fetal supporttissue product is obtained from a human, non-primate human, cow or pig.In some embodiments, the composition further comprises apharmaceutically-acceptable diluents, excipient or carrier. In someembodiments, the composition is formulated as a solution, suspension,paste, ointment, oil, emulsion, microemulsion, cream, lotion, gel, orcombination thereof. In some embodiments, the composition comprisessubstantially-isolated tissue product formulated into microspheres,microparticles, or liposomes. In some embodiments, the composition isformulated into PLGA copolymers. In some embodiments, the composition isformulated for controlled, sustained, or delayed release. In someembodiments, the composition is administered by a patch. In someembodiments, the patch is placed on an osteolytic bone or an osteolyticjoint. In some embodiments, the composition is administered by animplant. In some embodiments, the implant is placed on an osteolyticbone or an osteolytic joint. In some embodiments, the composition isformulated into an orthopaedic prosthesis. In some embodiments, thefetal support tissue product is in the form of a substantially-flattenedsheet. In some embodiments, the composition further comprises a backing.In some embodiments, the composition is wrapped around an osteolyticbone or an osteolytic joint. In some embodiments, the methods furthercomprise administering to the individual a calcium supplement. In someembodiments, the methods further comprise administering to theindividual a bisphosphonate.

Disclosed herein, in certain embodiments, are methods of treating a bonetumor, the methods comprising administering to an individual in needthereof a therapeutically-effective amount of a composition comprising afetal support tissue product or an extract thereof. In some embodiments,the fetal support tissue product is substantially isolated placentalamniotic membrane (PAM), umbilical cord amniotic membrane (UCAM),placenta, umbilical cord, chorion, amnion-chorion, or any combinationsthereof. In some embodiments, the fetal support tissue product is frozenor previously frozen PAM, frozen or previously frozen UCAM, frozen orpreviously frozen placenta, frozen or previously frozen umbilical cord,frozen or previously frozen chorion, frozen or previously frozenamnion-chorion, or any combinations thereof. In some embodiments, thefetal support tissue product is cryopreserved, lyophilized, terminallysterilized, or a combination thereof. In some embodiments, the fetalsupport tissue product is in the form of a pulverized powder or ahomogenate.tissue producttissue producttissue producttissue product Insome embodiments, the fetal support tissue product does not comprise avein or an artery, a cell with metabolic activity, active HIV-1, activeHIV-2, active HTLV-1, active hepatitis B, active hepatitis C, activeWest Nile Virus, active cytomegalovirus, active human transmissiblespongiform encephalopathy, or active treponema pallidum. In someembodiments, the fetal support tissue product is obtained from a human,non-primate human, cow or pig. In some embodiments, the compositionfurther comprises a pharmaceutically-acceptable diluents, excipient orcarrier. In some embodiments, the composition is formulated as asolution, suspension, paste, ointment, oil, emulsion, microemulsion,cream, lotion, gel, or combination thereof. In some embodiments, thecomposition comprises substantially-isolated tissue product formulatedinto microspheres, microparticles, or liposomes. In some embodiments,the composition is formulated into PLGA copolymers. In some embodiments,the composition is formulated for controlled, sustained, or delayedrelease. In some embodiments, the composition is injected into anosteolytic joint. In some embodiments, the composition is administeredby a patch. In some embodiments, the composition is administered by apatch. In some embodiments, the patch is placed on an osteolytic bone oran osteolytic joint. In some embodiments, the composition isadministered by an implant. In some embodiments, the implant is placedon an osteolytic bone or an osteolytic joint. In some embodiments, thecomposition is formulated into an orthopaedic prosthesis. In someembodiments, the fetal support tissue product is in the form of asubstantially-flattened sheet. In some embodiments, the compositionfurther comprises a backing. In some embodiments, the composition iswrapped around an osteolytic bone or an osteolytic joint. In someembodiments, the methods further comprise administering to theindividual a calcium supplement. In some embodiments, the methodsfurther comprise administering to the individual a chemotherapeuticagent, a bisphosphinate, Metastron, or any combination thereof.

Disclosed herein, in certain embodiments, are methods of inhibitingosteoclast differentiation in an individual in need thereof, comprisingadministering to the individual a composition comprising substantiallyisolated HC-HA complex. In some embodiments, the HC-HA complex isobtained by a process comprising: (a) providing a reaction mixturecomprising: (i) HA (e.g., HMW HA); (ii) IαI, wherein the IαI isoptionally in serum or isolated from serum; (iii); (iii) TSG-6, whereinthe TSG-6 is optionally recombinant; and (iv) PTX3, wherein the PTX3 isoptionally recombinant; wherein at least one of HA, IαI, TSG-6, or PTX3is optionally generated by a plurality of cells present in the reactionmixture; (b) incubating the reaction mixture for a period of timesufficient to produce HC-HA complex; and (c) isolating and purifying theHC-HA complex. In some embodiments, the substantially isolated HC-HAcomplex is derived from placental amniotic membrane (PAM), umbilicalcord amniotic membrane (UCAM), placenta, umbilical cord, chorion oramnion-chorion, and is optionally biochemically purified. In someembodiments, the HC-HA is purified by ultracentrifugation (e.g., fourrounds of ultracentrifugation). In some embodiments, the substantiallyisolated HC-HA complex is derived from frozen or previously frozenplacental amniotic membrane (PAM), frozen or previously frozen umbilicalcord amniotic membrane (UCAM), frozen or previously frozen placenta,frozen or previously frozen umbilical cord, frozen or previously frozenchorion or frozen or previously frozen amnion-chorion, and is optionallybiochemically purified. In some embodiments, the HC-HA is purified byultracentrifugation (e.g., four rounds of ultracentrifugation). In someembodiments, the composition further comprises apharmaceutically-acceptable diluents, excipient or carrier. In someembodiments, the composition is formulated as a solution, suspension,paste, ointment, oil, emulsion, microemulsion, cream, lotion, gel, orcombination thereof. In some embodiments, the composition comprisessubstantially-isolated tissue product formulated into microspheres,microparticles, or liposomes. In some embodiments, the composition isformulated into PLGA copolymers. In some embodiments, the composition isformulated for controlled, sustained, or delayed release. In someembodiments, the composition is injected into an osteolytic joint. Insome embodiments, the composition is administered by a patch. In someembodiments, the patch is placed on an osteolytic bone or an osteolyticjoint. In some embodiments, the composition is administered by animplant. In some embodiments, the implant is placed on an osteolyticbone or an osteolytic joint. In some embodiments, the composition isformulated into an orthopaedic prosthesis. In some embodiments, themethods further comprise administering to the individual a calciumsupplement.

Disclosed herein, in certain embodiments, are methods of promotingmineralization by osteoblasts in an individual in need thereof,comprising administering to the individual a composition comprisingsubstantially isolated HC-HA complex. In some embodiments, the HC-HAcomplex is obtained by a process comprising: (a) providing a reactionmixture comprising: (i) HA (e.g., HMW HA); (ii) IαI, wherein the IαI isoptionally in serum or isolated from serum; (iii); (iii) TSG-6; whereinthe TSG-6 is optionally recombinant; and (iv) PTX3, wherein the PTX3 isoptionally recombinant; wherein at least one of HA, IαI, TSG-6, TSG-6like protein is optionally generated by a plurality of cells present inthe reaction mixture; (b) incubating the reaction mixture for a periodof time sufficient to produce HC-HA complex; and (c) isolating andpurifying the HC-HA complex. In some embodiments, the substantiallyisolated HC-HA complex is derived from placental amniotic membrane(PAM), umbilical cord amniotic membrane (UCAM), placenta, umbilicalcord, chorion or amnion-chorion, and is optionally biochemicallypurified. In some embodiments, the substantially isolated HC-HA complexis derived from frozen or previously frozen placental amniotic membrane(PAM), frozen or previously frozen umbilical cord amniotic membrane(UCAM), frozen or previously frozen placenta, frozen or previouslyfrozen umbilical cord, frozen or previously frozen chorion, or frozen orpreviously frozen amnion-chorion, and is optionally biochemicallypurified. In some embodiments, the HC-HA is purified byultracentrifugation (e.g., four rounds of ultracentrifugation). In someembodiments, the composition further comprises apharmaceutically-acceptable diluents, excipient or carrier. In someembodiments, the composition is formulated as a solution, suspension,paste, ointment, oil, emulsion, microemulsion, cream, lotion, gel, orcombination thereof. In some embodiments, the composition comprisessubstantially-isolated tissue product formulated into microspheres,microparticles, or liposomes. In some embodiments, the composition isformulated into PLGA copolymers. In some embodiments, the composition isformulated for controlled, sustained, or delayed release. In someembodiments, the composition is injected into an osteolytic joint. Insome embodiments, the composition is administered by a patch. In someembodiments, the patch is placed on an osteolytic bone or an osteolyticjoint. In some embodiments, the composition is administered by animplant. In some embodiments, the implant is placed on an osteolyticbone or an osteolytic joint. In some embodiments, the composition isformulated into an orthopaedic prosthesis. In some embodiments, themethods further comprise administering to the individual a calciumsupplement.

Disclosed herein, in certain embodiments, are methods of inhibiting boneresorption in an individual in need thereof, comprising administering tothe individual a composition comprising substantially isolated HC-HAcomplex. In some embodiments, the HC-HA complex is obtained by a processcomprising: (a) providing a reaction mixture comprising: (i) HA (e.g.,HMW HA); (ii) IαI, wherein the IαI is optionally in serum or isolatedfrom serum; (iii); (iii) TSG-6, wherein the TSG-6 is optionallyrecombinant; and (iv) PTX3, wherein the PTX3 is optionally recombinant;wherein at least one of HA, IαI, TSG-6, or PTX3 is optionally generatedby a plurality of cells present in the reaction mixture; (b) incubatingthe reaction mixture for a period of time sufficient to produce HC-HAcomplex; and (c) isolating and purifying the HC-HA complex. In someembodiments, the substantially isolated HC-HA complex is derived fromplacental amniotic membrane (PAM), umbilical cord amniotic membrane(UCAM), placenta, umbilical cord, chorion or amnion-chorion, and isoptionally biochemically purified. In some embodiments, thesubstantially isolated HC-HA complex is derived from frozen orpreviously frozen placental amniotic membrane (PAM), frozen orpreviously frozen umbilical cord amniotic membrane (UCAM), frozen orpreviously frozen placenta, frozen or previously frozen umbilical cord,frozen or previously frozen chorion or frozen or previously frozenamnion-chorion, and is optionally biochemically purified. In someembodiments, the HC-HA is purified by ultracentrifugation (e.g., fourrounds of ultracentrifugation). In some embodiments, the compositionfurther comprises a pharmaceutically-acceptable diluents, excipient orcarrier. In some embodiments, the composition is formulated as asolution, suspension, paste, ointment, oil, emulsion, microemulsion,cream, lotion, gel, or combination thereof. In some embodiments, thecomposition comprises substantially-isolated tissue product formulatedinto microspheres, microparticles, or liposomes. In some embodiments,the composition is formulated into PLGA copolymers. In some embodiments,the composition is formulated for controlled, sustained, or delayedrelease. In some embodiments, the composition is injected into anosteolytic joint. In some embodiments, the composition is administeredby a patch. In some embodiments, the patch is placed on an osteolyticbone or an osteolytic joint. In some embodiments, the composition isadministered by an implant. In some embodiments, the implant is placedon an osteolytic bone or an osteolytic joint. In some embodiments, thecomposition is formulated into an orthopaedic prosthesis. In someembodiments, the methods further comprise administering to theindividual a calcium supplement.

Disclosed herein, in certain embodiments, are methods of inhibiting boneremodeling in an individual in need thereof, comprising administering tothe individual a composition comprising substantially isolated HC-HAcomplex. In some embodiments, the HC-HA complex is obtained by a processcomprising: (a) providing a reaction mixture comprising: (i) HA (e.g.,HMW HA); (ii) IαI, wherein the IαI is optionally in serum or isolatedfrom serum; (iii) TSG-6, wherein the TSG-6 is optionally recombinant;and (iv) PTX3, wherein the PTX3 is optionally recombinant; wherein atleast one of HA, IαI, TSG-6, or PTX3 is optionally generated by aplurality of cells present in the reaction mixture; (b) incubating thereaction mixture for a period of time sufficient to produce HC-HAcomplex; and (c) isolating and purifying the HC-HA complex. In someembodiments, the substantially isolated HC-HA complex is derived fromplacental amniotic membrane (PAM), umbilical cord amniotic membrane(UCAM), placenta, umbilical cord, chorion or amnion-chorion. In someembodiments, the substantially isolated HC-HA complex is derived fromfrozen or previously frozen placental amniotic membrane (PAM), frozen orpreviously frozen umbilical cord amniotic membrane (UCAM), frozen orpreviously frozen placenta, frozen or previously frozen umbilical cord,frozen or previously frozen chorion or frozen or previously frozenamnion-chorion. In some embodiments, the composition further comprises apharmaceutically-acceptable diluents, excipient or carrier. In someembodiments, the composition is formulated as a solution, suspension,paste, ointment, oil, emulsion, microemulsion, cream, lotion, gel, orcombination thereof. In some embodiments, the composition comprisessubstantially-isolated tissue product formulated into microspheres,microparticles, or liposomes. In some embodiments, the composition isformulated into PLGA copolymers. In some embodiments, the composition isformulated for controlled, sustained, or delayed release. In someembodiments, the composition is injected into an osteolytic joint. Insome embodiments, the composition is administered by a patch. In someembodiments, the patch is placed on an osteolytic bone or an osteolyticjoint. In some embodiments, the composition is administered by animplant. In some embodiments, the implant is placed on an osteolyticbone or an osteolytic joint. In some embodiments, the composition isformulated into an orthopaedic prosthesis. In some embodiments, themethods further comprise administering to the individual a calciumsupplement.

Disclosed herein, in certain embodiments, are methods of balancing boneresorption and bone formation, comprising administering to an individualin need thereof a therapeutically-effective amount of a compositioncomprising substantially isolated HC-HA complex. In some embodiments,the HC-HA complex is obtained by a process comprising: (a) providing areaction mixture comprising: (i) HA (e.g., HMW HA); (ii) IαI, whereinthe IαI is optionally in serum or isolated from serum; (iii) TSG-6,wherein the TSG-6 is optionally recombinant; and (iv) PTX3, wherein thePTX3 is optionally recombinant; wherein at least one of HA, IαI, TSG-6,or PTX3 is optionally generated by a plurality of cells present in thereaction mixture; (b) incubating the reaction mixture for a period oftime sufficient to produce HC-HA complex; and (c) isolating andpurifying the HC-HA complex. In some embodiments, the substantiallyisolated HC-HA complex is derived from placental amniotic membrane(PAM), umbilical cord amniotic membrane (UCAM), placenta, umbilicalcord, chorion or amnion-chorion, and is optionally biochemicallypurified. In some embodiments, the substantially isolated HC-HA complexis derived from frozen or previously frozen placental amniotic membrane(PAM), frozen or previously frozen umbilical cord amniotic membrane(UCAM), frozen or previously frozen placenta, frozen or previouslyfrozen umbilical cord, frozen or previously frozen chorion or frozen orpreviously frozen amnion-chorion, and is optionally biochemicallypurified. In some embodiments, the HC-HA is purified byultracentrifugation (e.g., four rounds of ultracentrifugation). In someembodiments, the composition further comprises apharmaceutically-acceptable diluents, excipient or carrier. In someembodiments, the composition is formulated as a solution, suspension,paste, ointment, oil, emulsion, microemulsion, cream, lotion, gel, orcombination thereof. In some embodiments, the composition comprisessubstantially-isolated tissue product formulated into microspheres,microparticles, or liposomes. In some embodiments, the composition isformulated into PLGA copolymers. In some embodiments, the composition isformulated for controlled, sustained, or delayed release. In someembodiments, the composition is injected into an osteolytic joint. Insome embodiments, the composition is administered by a patch. In someembodiments, the patch is placed on an osteolytic bone or an osteolyticjoint. In some embodiments, the composition is administered by animplant. In some embodiments, the implant is placed on an osteolyticbone or an osteolytic joint. In some embodiments, the composition isformulated into an orthopaedic prosthesis. In some embodiments, themethods further comprise administering to the individual a calciumsupplement.

Disclosed herein, in certain embodiments, are methods of treating adisease, disorder, or condition characterized by excessive or undesiredosteoclast differentiation, the methods comprising administering to anindividual in need thereof a therapeutically-effective amount of acomposition comprising substantially isolated HC-HA complex. In someembodiments, the HC-HA complex is obtained by a process comprising: (a)providing a reaction mixture comprising: (i) HA (e.g., HMW HA); (ii)IαI, wherein the IαI is optionally in serum or isolated from serum;(iii); (iii) TSG-6, wherein the TSG-6 is optionally recombinant; and(iv) PTX3, wherein the PTX3 is optionally recombinant; wherein at leastone of HA, IαI, TSG-6, or PTX3 is optionally generated by a plurality ofcells present in the reaction mixture; (b) incubating the reactionmixture for a period of time sufficient to produce HC-HA complex; and(c) isolating and purifying the HC-HA complex. In some embodiments, thesubstantially isolated HC-HA complex is derived from placental amnioticmembrane (PAM), umbilical cord amniotic membrane (UCAM), placenta,umbilical cord, chorion or amnion-chorion, and is optionallybiochemically purified. In some embodiments, the substantially isolatedHC-HA complex is derived from frozen or previously frozen placentalamniotic membrane (PAM), frozen or previously frozen umbilical cordamniotic membrane (UCAM), frozen or previously frozen placenta, frozenor previously frozen umbilical cord, frozen or previously frozen chorionor frozen or previously frozen amnion-chorion, and is optionallybiochemically purified. In some embodiments, the HC-HA is purified byultracentrifugation (e.g., four rounds of ultracentrifugation). In someembodiments, the composition further comprises apharmaceutically-acceptable diluents, excipient or carrier. In someembodiments, the composition is formulated as a solution, suspension,paste, ointment, oil, emulsion, microemulsion, cream, lotion, gel, orcombination thereof. In some embodiments, the composition comprisessubstantially-isolated tissue product formulated into microspheres,microparticles, or liposomes. In some embodiments, the composition isformulated into PLGA copolymers. In some embodiments, the composition isformulated for controlled, sustained, or delayed release. In someembodiments, the composition is injected into an osteolytic joint. Insome embodiments, the composition is administered by a patch. In someembodiments, the patch is placed on an osteolytic bone or an osteolyticjoint. In some embodiments, the composition is administered by animplant. In some embodiments, the implant is placed on an osteolyticbone or an osteolytic joint. In some embodiments, the composition isformulated into an orthopaedic prosthesis. In some embodiments, themethods further comprise administering to the individual a calciumsupplement.

Disclosed herein, in certain embodiments, are methods of treating adisease, disorder, or condition characterized by deficient or defectivebone formation, the methods comprising administering to an individual inneed thereof a therapeutically-effective amount of a compositioncomprising substantially isolated HC-HA complex. In some embodiments,the HC-HA complex is obtained by a process comprising: (a) providing areaction mixture comprising: (i) HA (e.g., HMW HA); (ii) IαI, whereinthe IαI is optionally in serum or isolated from serum; (iii); (iii)TSG-6, wherein the TSG-6 is optionally recombinant; and (iv) PTX3,wherein the PTX3 is optionally recombinant; wherein at least one of HA,IαI, TSG-6, or PTX3 is optionally generated by a plurality of cellspresent in the reaction mixture; (b) incubating the reaction mixture fora period of time sufficient to produce HC-HA complex; and (c) isolatingand purifying the HC-HA complex. In some embodiments, the substantiallyisolated HC-HA complex is derived from placental amniotic membrane(PAM), umbilical cord amniotic membrane (UCAM), placenta, umbilicalcord, chorion or amnion-chorion, and is optionally biochemicallypurified. In some embodiments, the substantially isolated HC-HA complexis derived from frozen or previously frozen placental amniotic membrane(PAM), frozen or previously frozen umbilical cord amniotic membrane(UCAM), frozen or previously frozen placenta, frozen or previouslyfrozen umbilical cord, frozen or previously frozen chorion or frozen orpreviously frozen amnion-chorion, and is optionally biochemicallypurified. In some embodiments, the HC-HA is purified byultracentrifugation (e.g., four rounds of ultracentrifugation). In someembodiments, the composition further comprises apharmaceutically-acceptable diluents, excipient or carrier. In someembodiments, the composition is formulated as a solution, suspension,paste, ointment, oil, emulsion, microemulsion, cream, lotion, gel, orcombination thereof. In some embodiments, the composition comprisessubstantially-isolated tissue product formulated into microspheres,microparticles, or liposomes. In some embodiments, the composition isformulated into PLGA copolymers. In some embodiments, the composition isformulated for controlled, sustained, or delayed release. In someembodiments, the composition is injected into an osteolytic joint. Insome embodiments, the composition is administered by a patch. In someembodiments, the patch is placed on an osteolytic bone or an osteolyticjoint. In some embodiments, the composition is administered by animplant. In some embodiments, the implant is placed on an osteolyticbone or an osteolytic joint. In some embodiments, the composition isformulated into an orthopaedic prosthesis. In some embodiments, themethods further comprise administering to the individual a calciumsupplement.

Disclosed herein, in certain embodiments, are methods of treatingarthritis, the methods comprising administering to an individual in needthereof a therapeutically-effective amount of a composition comprisingsubstantially isolated HC-HA complex. In some embodiments, the arthritisis osteoarthritis, rheumatoid arthritis, psoriatic arthritis, or anycombination thereof. In some embodiments, the HC-HA complex is obtainedby a process comprising: (a) providing a reaction mixture comprising:(i) HA (e.g., HMW HA); (ii) IαI, wherein the IαI is optionally in serumor isolated from serum; (iii); (iii) TSG-6, wherein the TSG-6 isoptionally recombinant; and (iv) PTX3, wherein the PTX3 is optionallyrecombinant; wherein at least one of HA, IαI, TSG-6, or PTX3 isoptionally generated by a plurality of cells present in the reactionmixture; (b) incubating the reaction mixture for a period of timesufficient to produce HC-HA complex; and (c) isolating and purifying theHC-HA complex. In some embodiments, the substantially isolated HC-HAcomplex is derived from placental amniotic membrane (PAM), umbilicalcord amniotic membrane (UCAM), placenta, umbilical cord, chorion oramnion-chorion, and is optionally biochemically purified. In someembodiments, the substantially isolated HC-HA complex is derived fromfrozen or previously frozen placental amniotic membrane (PAM), frozen orpreviously frozen umbilical cord amniotic membrane (UCAM), frozen orpreviously frozen placenta, frozen or previously frozen umbilical cord,frozen or previously frozen chorion or frozen or previously frozenamnion-chorion, and is optionally biochemically purified. In someembodiments, the HC-HA is purified by ultracentrifugation (e.g., fourrounds of ultracentrifugation). In some embodiments, the compositionfurther comprises a pharmaceutically-acceptable diluents, excipient orcarrier. In some embodiments, the composition is formulated as asolution, suspension, paste, ointment, oil, emulsion, microemulsion,cream, lotion, gel, or combination thereof. In some embodiments, thecomposition comprises substantially-isolated tissue product formulatedinto microspheres, microparticles, or liposomes. In some embodiments,the composition is formulated into PLGA copolymers. In some embodiments,the composition is formulated for controlled, sustained, or delayedrelease. In some embodiments, the composition is injected into anosteolytic joint. In some embodiments, the composition is administeredby a patch. In some embodiments, the patch is placed on an osteolyticbone or an osteolytic joint. In some embodiments, the composition isadministered by an implant. In some embodiments, the implant is placedon an osteolytic bone or an osteolytic joint. In some embodiments, thecomposition is formulated into an orthopaedic prosthesis. In someembodiments, the methods further comprise administering to theindividual a calcium supplement. In some embodiments, the methodsfurther comprise administering to the individual an NSAID, acorticosteroid, hyaluronan injections, a DMARD, an analgesic, or anycombination thereof.

Disclosed herein, in certain embodiments, are methods of treatingosteoporosis, the methods comprising administering to an individual inneed thereof a therapeutically-effective amount of a compositioncomprising substantially isolated HC-HA complex. In some embodiments,the HC-HA complex is obtained by a process comprising: (a) providing areaction mixture comprising: (i) HA (e.g., HMW HA); (ii) IαI, whereinthe IαI is optionally in serum or isolated from serum; (iii); (iii)TSG-6, wherein the TSG-6 is optionally recombinant; and (iv) PTX3,wherein the PTX3 is optionally recombinant; wherein at least one of HA,IαI, TSG-6, or PTX3 is optionally generated by a plurality of cellspresent in the reaction mixture; (b) incubating the reaction mixture fora period of time sufficient to produce HC-HA complex; and (c) isolatingand purifying the HC-HA complex. In some embodiments, the substantiallyisolated HC-HA complex is derived from placental amniotic membrane(PAM), umbilical cord amniotic membrane (UCAM), placenta, umbilicalcord, chorion or amnion-chorion, and is optionally biochemicallypurified. In some embodiments, the substantially isolated HC-HA complexis derived from frozen or previously frozen placental amniotic membrane(PAM), frozen or previously frozen umbilical cord amniotic membrane(UCAM), frozen or previously frozen placenta, frozen or previouslyfrozen umbilical cord, frozen or previously frozen chorion or frozen orpreviously frozen amnion-chorion, and is optionally biochemicallypurified. In some embodiments, the HC-HA is purified byultracentrifugation (e.g., four rounds of ultracentrifugation). In someembodiments, the composition further comprises apharmaceutically-acceptable diluents, excipient or carrier. In someembodiments, the composition is formulated as a solution, suspension,paste, ointment, oil, emulsion, microemulsion, cream, lotion, gel, orcombination thereof. In some embodiments, the composition comprisessubstantially-isolated tissue product formulated into microspheres,microparticles, or liposomes. In some embodiments, the composition isformulated into PLGA copolymers. In some embodiments, the composition isformulated for controlled, sustained, or delayed release. In someembodiments, the composition is injected into an osteolytic joint. Insome embodiments, the composition is administered by a patch. In someembodiments, the patch is placed on an osteolytic bone or an osteolyticjoint. In some embodiments, the composition is administered by animplant. In some embodiments, the implant is placed on an osteolyticbone or an osteolytic joint. In some embodiments, the composition isformulated into an orthopaedic prosthesis. In some embodiments, themethods further comprise administering to the individual a calciumsupplement. In some embodiments, the methods further compriseadministering to the individual a bisphosphonate, an estrogen analog,Raloxifene, Calcitonin, Teriparatide, calcium salts, sodium fluoride,RANKL inhibitors, Strontium ranelate, or any combination thereof.

Disclosed herein, in certain embodiments, are methods of treatingalveolar bone degradation, the methods comprising administering to anindividual in need thereof a therapeutically-effective amount of acomposition comprising substantially isolated HC-HA complex. In someembodiments, the HC-HA complex is obtained by a process comprising: (a)providing a reaction mixture comprising: (i) HA (e.g., HMW HA); (ii)IαI, wherein the IαI is optionally in serum or isolated from serum;(iii) TSG-6, wherein the TSG-6 is optionally recombinant; and (iv) PTX3,wherein the PTX3 is optionally recombinant; wherein at least one of HA,IαI, TSG-6, or PTX3 is optionally generated by a plurality of cellspresent in the reaction mixture; (b) incubating the reaction mixture fora period of time sufficient to produce HC-HA complex; and (c) isolatingand purifying the HC-HA complex. In some embodiments, the substantiallyisolated HC-HA complex is derived from placental amniotic membrane(PAM), umbilical cord amniotic membrane (UCAM), placenta, umbilicalcord, chorion or amnion-chorion, and is optionally biochemicallypurified. In some embodiments, the substantially isolated HC-HA complexis derived from frozen or previously frozen placental amniotic membrane(PAM), frozen or previously frozen umbilical cord amniotic membrane(UCAM), frozen or previously frozen placenta, frozen or previouslyfrozen umbilical cord, frozen or previously frozen chorion or frozen orpreviously frozen amnion-chorion, and is optionally biochemicallypurified. In some embodiments, the HC-HA is purified byultracentrifugation (e.g., four rounds of ultracentrifugation). In someembodiments, the composition further comprises apharmaceutically-acceptable diluents, excipient or carrier. In someembodiments, the composition is formulated as a solution, suspension,paste, ointment, oil, emulsion, microemulsion, cream, lotion, gel, orcombination thereof. In some embodiments, the composition comprisessubstantially-isolated tissue product formulated into microspheres,microparticles, or liposomes. In some embodiments, the composition isformulated into PLGA copolymers. In some embodiments, the composition isformulated for controlled, sustained, or delayed release. In someembodiments, the composition is administered by a patch. In someembodiments, the patch is placed on the alveolar bone. In someembodiments, the composition is administered by an implant. In someembodiments, the implant is placed on an alveolar bone. In someembodiments, the methods further comprise administering to theindividual a calcium supplement.

Disclosed herein, in certain embodiments, are methods of treatingPaget's disease, the methods comprising administering to an individualin need thereof a therapeutically-effective amount of a compositioncomprising substantially isolated HC-HA complex. In some embodiments,the HC-HA complex is obtained by a process comprising: (a) providing areaction mixture comprising: (i) HA (e.g., HMW HA); (ii) IαI, whereinthe IαI is optionally in serum or isolated from serum; (iii) TSG-6,wherein the TSG-6 is optionally recombinant; and (iv) PTX3, wherein thePTX3 is optionally recombinant; wherein at least one of HA, IαI, TSG-6,or PTX3 is optionally generated by a plurality of cells present in thereaction mixture; (b) incubating the reaction mixture for a period oftime sufficient to produce HC-HA complex; and (c) isolating andpurifying the HC-HA complex. In some embodiments, the substantiallyisolated HC-HA complex is derived from placental amniotic membrane(PAM), umbilical cord amniotic membrane (UCAM), placenta, umbilicalcord, chorion or amnion-chorion, and is optionally biochemicallypurified. In some embodiments, the substantially isolated HC-HA complexis derived from frozen or previously frozen placental amniotic membrane(PAM), frozen or previously frozen umbilical cord amniotic membrane(UCAM), frozen or previously frozen placenta, frozen or previouslyfrozen umbilical cord, frozen or previously frozen chorion or frozen orpreviously frozen amnion-chorion, and is optionally biochemicallypurified. In some embodiments, the HC-HA is purified byultracentrifugation (e.g., four rounds of ultracentrifugation). In someembodiments, the composition further comprises apharmaceutically-acceptable diluents, excipient or carrier. In someembodiments, the composition is formulated as a solution, suspension,paste, ointment, oil, emulsion, microemulsion, cream, lotion, gel, orcombination thereof. In some embodiments, the composition comprisessubstantially-isolated tissue product formulated into microspheres,microparticles, or liposomes. In some embodiments, the composition isformulated into PLGA copolymers. In some embodiments, the composition isformulated for controlled, sustained, or delayed release. In someembodiments, the composition is injected into an osteolytic joint. Insome embodiments, the composition is administered by a patch. In someembodiments, the patch is placed on an osteolytic bone or an osteolyticjoint. In some embodiments, the composition is administered by animplant. In some embodiments, the implant is placed on an osteolyticbone or an osteolytic joint. In some embodiments, the composition isformulated into an orthopaedic prosthesis. In some embodiments, themethods further comprise administering to the individual a calciumsupplement. In some embodiments, the methods further compriseadministering to the individual a bisphosphonate.

Disclosed herein, in certain embodiments, are methods of treating a bonetumor, the methods comprising administering to an individual in needthereof a therapeutically-effective amount of a composition comprisingsubstantially isolated HC-HA complex. In some embodiments, the HC-HAcomplex is obtained by a process comprising: (a) providing a reactionmixture comprising: (i) HA (e.g., HMW HA); (ii) IαI, wherein the IαI isoptionally in serum or isolated from serum; (iii) TSG-6, wherein theTSG-6 is optionally recombinant; and (iv) PTX3, wherein the PTX3 isoptionally recombinant; wherein at least one of HA, IαI, TSG-6, or PTX3is optionally generated by a plurality of cells present in the reactionmixture; (b) incubating the reaction mixture for a period of timesufficient to produce HC-HA complex; and (c) isolating and purifying theHC-HA complex. In some embodiments, the substantially isolated HC-HAcomplex is derived from placental amniotic membrane (PAM), umbilicalcord amniotic membrane (UCAM), placenta, umbilical cord, chorion oramnion-chorion, and is optionally biochemically purified. In someembodiments, the substantially isolated HC-HA complex is derived fromfrozen or previously frozen placental amniotic membrane (PAM), frozen orpreviously frozen umbilical cord amniotic membrane (UCAM), frozen orpreviously frozen placenta, frozen or previously frozen umbilical cord,frozen or previously frozen chorion or frozen or previously frozenamnion-chorion, and is optionally biochemically purified. In someembodiments, the HC-HA is purified by ultracentrifugation (e.g., fourrounds of ultracentrifugation). In some embodiments, the compositionfurther comprises a pharmaceutically-acceptable diluents, excipient orcarrier. In some embodiments, the composition is formulated as asolution, suspension, paste, ointment, oil, emulsion, microemulsion,cream, lotion, gel, or combination thereof. In some embodiments, thecomposition comprises substantially-isolated tissue product formulatedinto microspheres, microparticles, or liposomes. In some embodiments,the composition is formulated into PLGA copolymers. In some embodiments,the composition is formulated for controlled, sustained, or delayedrelease. In some embodiments, the composition is injected into anosteolytic joint. In some embodiments, the composition is administeredby a patch. In some embodiments, the patch is placed on an osteolyticbone or an osteolytic joint. In some embodiments, the composition isadministered by an implant. In some embodiments, the implant is placedon an osteolytic bone or an osteolytic joint. In some embodiments, thecomposition is formulated into an orthopaedic prosthesis. In someembodiments, the methods further comprise administering to theindividual a calcium supplement. In some embodiments, the methodsfurther comprise administering to the individual a chemotherapeuticagent, a bisphosphinate, Metastron, or any combination thereof.

BRIEF DESCRIPTION OF THE DRAWINGS

The novel features of the invention are set forth with particularity inthe appended claims. A better understanding of the features andadvantages of the present invention will be obtained by reference to thefollowing detailed description that sets forth illustrative embodiments,in which the principles of the invention are utilized, and theaccompanying drawings of which:

FIG. 1 exemplifies the formation of osteoclast is regulated byRANK/RANKL and its downstream signaling pathway. NFATc1 is a criticaltranscriptional factor that regulates the expression ofosteoclastogenesis-related genes including TRAP.

FIG. 2 exemplifies macrophage RAW 264.7 cells cultured with (E-G, PosCTL) and without (A-C, Neg CTL) 50 ng/ml RANKL stimulation. Osteoclastsbegan to form on Day 3 (F) and complete differentiation was observed onDay 5 (G) as evidenced by positive TRAP staining Bar=200 μm.

FIG. 3 exemplifies macrophage RAW 264.7 cells stimulated with 50 ng/mlRANKL and cultured with soluble amniotic membrane extract (AME) (A),HC-HA complex purified from AM (HC-HA) (B), AM lysate (AML) (C), AMpowder (AMP) (D), and on the epithelial side of intact amniotic membrane(iAM)(E), the stromal side of iAM (F), the basement membrane side ofepithelially-denuded AM (dAM) (G), the stromal side of dAM (H), theepithelial side of lyophilized amniotic membrane lAM (I), or the stromalside of lyophilized amniotic membrane lAM (J). Cell morphology on Day 3(Column 1), Day 5 (A2-D2), and Day 6 (E2-J2). TRAP staining (Column 3)revealed no large multinucleated osteoclasts when cultured on AM and itsderivatives. Bar=200 μm.

FIG. 4 exemplifies Neg CTL showing significantly lower (p=0.001) TRAPreading compared to the Pos CTL. Inhibitory action for osteoclastformation was seen on all AM derivatives (200 μg/ml protein) with HC-HA(25 μg/ml HA) being the most potent followed by AME, AMP, and AML.

FIG. 6 exemplifies Neg CTL showing significantly lower (p=2.9E-12) TRAPreading compared to Pos CTL. Statistically significant inhibitory actionfor osteoclast formation (all p<0.05) was seen on all AM tissuesincluding epithelial side of iAM, stromal side of iAM, basement membraneside of dAM and stromal side of dAM. Inhibitory action of osteoclast wasalso preserved after gamma irradiation of all AM tissues.

FIG. 5 exemplifies mRNA expression of NFATc1 (measured by quantitativePCR on Day 5 as being significantly downregulated by all AM derivatives(p<0.05).

FIG. 7 exemplifies mRNA expression of RANK on Day 7 as beingsignificantly downregulated for both HC-HA and stromal side of dAM.

FIG. 8 exemplifies expression of β3-integrin, which is required fornormal osteoclast differentiation, as being significantly downregulatedby HC-HA, AML, AMP, and stromal side of dAM. Suppression of β3-integrinby AME is statistically significant on Day 5 (p=0.000001).

FIG. 9 exemplifies the effects of nHC-HA on osteoclastogenesis. RAW264.7cells is effectively induced into osteoclasts at Day 6 with 50 ng/mlRANKL(a). The inhibition of osteoclast formation is dose dependent onnHC-HA purified from AM. (HC-HA) concentrations inhibit osteoclastformation while the highest concentration of high molecular weight HA(100 μg/ml) does not inhibit the osteoclast formation (b). The colorpicture of TRAP colorimetric assay is provided. The lighter of thecolor, the more inhibition of the TRAP activity is (c). nHC-HA as low as0.08 μg/ml significantly inhibits the osteoclast formation (d). IC50 ofHC-HA on the osteoclast formation is calculated to be about 0.1 μg/ml(e).

FIG. 10 exemplifies the effects of AMP (amniotic membrane powder) onosteoclastogenesis. Previous results can be reliably reproduced (a).Like HC-HA, AMP dose (0-500 μg/ml protein) dependently inhibits theosteoclast formation (b). The color picture of TRAP colorimetric assayis provided (c and d). Osteoclast inhibition by AMP is measured by TRAPColorimetric assay (e). IC50 of AMP on the osteoclast formation iscalculated to be about 20.1 μg/ml (f).

FIG. 11 exemplifies the performance of several tissue products (AME,HC-HA, AML, AMP) in inhibiting osteoclast formation. All AM derivatessignificantly inhibit osteoclast formation, and AMP prepared fromdifferent donors (AMP 1-5) consistently showed inhibitory activity thatwere more potent than AML.

FIG. 12 exemplifies the performance of AMP in inhibiting osteoclastformation. Powders were made from amniotic membrane (AMP), chorion(CHP), amnio-chorion (ACP), whole placenta (PLP), whole umbilical cord(UCP), and umbilical cord amniotic membane (UAMP). Like HC-HA (5 μg/ml),AMP (200 μg/ml), ACP (100 μg/ml), PLP (100 μg/ml), and WUC (100 μg/ml)significantly inhibit osteoclast formation.

FIG. 13 exemplifies the effects of AMP on osteogenesis. Aftercultivation for 7 days, the cell morphology of groups with induction(Pos Ctrl, HC-HA, and AMP) is changed: a compact layer of cells appearsin the periphery in Pos Ctrl and HC-HA, but becomes clusters in AMP.Alizarin staining (ARS) shows only the periphery is stained in Pos Ctrland HC-HA, bur both the periphery and the center are stained in AMP.

FIG. 14 exemplifies the effects of AMP on bone matrix mineralization.When ARS is measured quantitatively, cell treated with AMP (125 μg/ml)significantly promotes the mineralization (p=0.0001).

FIG. 15 exemplifies the effects of AMP on the proliferation ofosteoblasts. The cell proliferation in the control is increased from Day1 to Day 4. In contrast, with the treatment of AMP (125 μg/ml), the cellproliferation is first decreased from Day 1 to Day 2, and then increasedfrom Day 2 to Day 4. However, compared to that of the control, the cellproliferation with AMP treatment is significantly lower at Day 2 and Day4 (p<0.05).

FIG. 16 exemplifies the effects of AMP and mAMP on osteoclastogenesis.mAMP is a powder of injectable AminoFix, which is prepared by MimedxGroup (Kennesaw, Ga.) using Purion^(SM) Process, which is different fromwhat is disclosed in this invention based on heat drying and otherchemical processing. Both AMP and mAMP are suspended into PBS andprepared at a series of concentrations (8, 40, and 200 μg protein/ml)and added to RANKL-induced RAW264.7 cells. After 5 days treatment, theTRAP activity in cells is measured. AMP significantly anddose-dependently inhibits TRAP activity (p<0.05). Additionally, AMP issignificantly more potent than mAMP at the same concentration (p<0.05).

The abbreviation AME means amniotic membrane extract. AME was made byextraction of amniotic membrane with PBS followed by centrifugation toremove the pellet.

The abbreviation AML means amniotic membrane lysate. AML was made byhomogenating the wet amniotic membrane without adding any buffers.

The abbreviation AMP means amniotic membrane powder. AMP was made bylyophilizing amniotic membrane followed by pulverizing it into powder.

DETAILED DESCRIPTION OF THE INVENTION Certain Definitions

As used herein, “placenta” means the organ that connects a developingfetus to the maternal uterine wall to allow nutrient uptake, wasteelimination, and gas exchange via the maternal blood supply. Theplacenta is composed of three layers. The innermost placental layersurrounding the fetus is called amnion. The allantois is the middlelayer of the placenta (derived from the embryonic hindgut); bloodvessels originating from the umbilicus traverse this membrane. Theoutermost layer of the placenta, the chorion, comes into contact withthe endometrium. The chorion and allantois fuse to form thechorioallantoic membrane.

As used herein, “chorion” means the membrane formed by extraembryonicmesoderm and the two layers of trophoblast. The chorionic villi emergefrom the chorion, invade the endometrium, and allow transfer ofnutrients from maternal blood to fetal blood. The chorion consists oftwo layers: an outer formed by the trophoblast, and an inner formed bythe somatic mesoderm; the amnion is in contact with the latter. Thetrophoblast is made up of an internal layer of cubical or prismaticcells, the cytotrophoblast or layer of Langhans, and an external layerof richly nucleated protoplasm devoid of cell boundaries, thesyncytiotrophoblast. The avascular amnion is adherent to the inner layerof the chorion.

As used herein, “amnion-chorion” means a product comprising amnion andchorion. In some embodiments, the amnion and the chorion are notserperated (i.e., the amnion is naturally adherent to the inner layer ofthe chorion). In some embodiments, the amnion is initially separatedfrom the chorion and later combined with the chorion during processing.

As used herein, “umbilical cord” means the organ that connects adeveloping fetus to the placenta. The umbilical cord is composed ofWharton's jelly, a gelatinous substance made largely frommucopolysaccharides. It contains one vein, which carries oxygenated,nutrient-rich blood to the fetus, and two arteries that carrydeoxygenated, nutrient-depleted blood away.

As used herein, “placental amniotic membrane” (PAM) means amnioticmembrane derived from the placenta. In some embodiments, the PAM issubstantially isolated.

As used herein, “umbilical cord amniotic membrane” (UCAM) means amnioticmembrane derived from the umbilical cord. UCAM is a translucentmembrane. The UCAM has multiple layers an epithelial layer, a basementmembrane; a compact layer; a fibroblast layer; and a spongy layer. Itlacks blood vessels or a direct blood supply. In some embodiments, theUCAM is substantially isolated. In some embodiments, the UCAM comprisesWharton's Jelly. In some embodiments, the UCAM comprises blood vesselsand/or arteries. In some embodiments, the UCAM comprises Wharton's Jellyand blood vessels and/or arteries.

As used herein, “human cells, tissues, or cellular or tissue-basedproducts (HCT/Ps)” means articles containing or consisting of humancells or tissues that are intended for implantation, transplantation,infusion, or transfer into a human recipient.

As used herein, “minimal manipulation” means (1) for structural tissue,processing that does not alter the original relevant characteristics ofthe tissue relating to the tissue's utility for reconstruction, repair,or replacement; and (2) for cells or nonstructural tissues, processingthat does not alter the relevant biological characteristics of cells ortissues.

As used herein, ‘homologous use” means the repair, reconstruction,replacement, or supplementation of a recipient's cells or tissues withan HCT/P that performs the same basic function or functions in therecipient as in the donor.

As used herein, “fetal support tissue product” means any productcomprising tissue that is supports the development of a fetus. Examplesof fetal support tissue include, but are not limited to, (i) placentalamniotic membrane (PAM), or substantially isolated PAM, (ii) umbilicalcord amniotic membrane (UCAM) or substantially isolated UCAM, (iii)chorion or substantially isolated chorion, (iv) amnion-chorion orsubstantially isolated amnion-chorion, (v) placenta or substantiallyisolated placenta, (vi) umbilical cord or substantially isolatedumbilical cord, or (vii) any combinations thereof. In some embodiments,the fetal support tissue product is a sheet, a powder, or homogenate.

As used herein, “HC-HA”, means the covalent complex formed between heavychains (HCs) of inter-alpha-inhibitor (IαI) and hyaluronan (HA) by thecatalytic action of tumor necrosis factor (TNF)-stimulated gene-6(TSG-6).

As used herein, “sheet” means any continuous expanse or surface. In someembodiments, a fetal support tissue product described herein is a flatsheet. The sheet can be any shape and size. In some embodiments, thesheet is a square, circle, triangle, or rectangle. In some embodiments,the sheet comprises multiple layers (e.g., of chorion, amnion-chorion,UCAM, PAM, placenta, umbilical cord or any combinations thereof).

As used herein, “homogenized” means (a fetal support tissue product thathas been broken up into particles that are of substantially uniformsize.

“Substantially isolated” or “isolated” means that the fetal supporttissue product has been separate from undesired materials (e.g., redblood cells, blood vessels, and arteries) derived from the originalsource organism. Purity, or “isolation” may be assayed by standardmethods, and will ordinarily be at least about 10% pure, more ordinarilyat least about 20% pure, generally at least about 30% pure, and moregenerally at least about 40% pure; in further embodiments at least about50% pure, or more often at least about 60% pure; in still otherembodiments, at least about 95% pure.

As used herein, the substantial preservation of biological activity orstructural integrity means that when compared to the biological activityand structural integrity of non-processed tissue, the biologicalactivity and structural integrity of the fetal support tissue producthas only decreased by about 5%, about 10%, about 15%, about 20%, about25%, about 30%, about 35%, about 40%, about 50%, about 60%.

The term “fresh” refers to tissue that is less than 10 days oldfollowing birth, and which is in substantially the same form as it wasfollowing birth.

As used herein, “biological activity” means the activity of polypeptidesand polysaccharides. In some embodiments, the activity of polypeptidesand polysaccharides found in umbilical cord (and substantially isolatedumbilical cord), UCAM (and substantially isolated UCAM), placenta (andsubstantially isolated placenta), PAM (and substantially isolated PAM),chorion (and substantially isolated chorion), or amnion-chorion (andsubstantially isolated amnion-chorion).

As used herein, “structural integrity” means the integrity of stroma andbasement membrane that make up the UCAM, chorion, or amnion-chorion. Insome embodiments, the structural integrity of the UCAM results in suturepull out strength.

As used herein, “hyaluronan” (or “HA”) means a substantiallynon-sulfated or non-sulfated glycosaminoglycan with linear repeatingdisaccharide units of glucuronosyl-N-acetylglucosamine. In someembodiments, HA is obtained from a commercial supplier (e.g., SigmaAldrich or Abbott Medical Optics, Irvine, Calif.). In some embodiments,HA is obtained from a commercial supplier as a powder. In someembodiments, HA is obtained from a cell that expresses a hyaluronansynthases (e.g., HAS1, HAS2, and HAS3). In certain instances, an HAsynthase lengthens hyaluronan by repeatedly adding glucuronic acid andN-acetylglucosamine to the nascent polysaccharide as it is extrudedthrough the cell membrane into the extracellular space.

As used herein, “recombinant TSG-6” means a TSG-6 protein that isproduced by recombinant methods (i.e., the TSG-6 gene from a firstsource (e.g., a human TSG-6 gene) is cloned into a DNA molecule from asecond source (e.g., a bacterial plasmid).

As used herein, “recombinant PTX3” means a PTX3 protein that is producedby recombinant methods (i.e., the PTX3 gene from a first source (e.g., ahuman PTX3 gene) is cloned into a DNA molecule from a second source(e.g., a bacterial plasmid).

As used herein, the “production bioreactor” is the bioreactor in whichthe final HC-HA complex disclosed herein is made.

The terms “subject” and “individual” are used interchangeably. As usedherein, both terms mean any animal, preferably a mammal, including ahuman or non-human. The terms patient, subject, and individual are usedinterchangeably. None of the terms are to be interpreted as requiringthe supervision of a medical professional (e.g., a doctor, nurse,physician's assistant, orderly, hospice worker).

The terms “treat,” “treating” or “treatment,” as used herein, includealleviating, abating or ameliorating a disease or condition symptoms,preventing additional symptoms, ameliorating or preventing theunderlying metabolic causes of symptoms, inhibiting the disease orcondition, e.g., arresting the development of the disease or condition,relieving the disease or condition, causing regression of the disease orcondition, relieving a condition caused by the disease or condition, orstopping the symptoms of the disease or condition eitherprophylactically and/or therapeutically.

Methods of Use

Bone Remodeling

Bone remodeling (or bone metabolism) is the process by which mature bonetissue is removed from the skeleton (bone resorption) and new bonetissue is formed (ossification).

Bone resorption is the process by which osteoclasts break down bone,releasing minerals from bone into the blood. Osteoclasts are largemultinucleated cells formed by the fusion of cells from themonocyte/macrophage cell line. Osteoclasts are generally present on theouter layer of bone, just beneath the periosteum. Attachment of theosteoclast to the osteon begins the process. The osteoclast then inducesan infolding of its cell membrane and secretes collagenase. Calcium,magnesium, phosphate and products of collagen are released into blood asthe osteoclasts tunnel into the mineralized bone. Osteoclasts have beenindicated in skeletal diseases such as osteoporosis, rheumatoidarthritis and Paget's disease, and are responsible for osteolysis.

Osteoclasts have been indicated in skeletal diseases such asosteoporosis, rheumatoid arthritis and Paget's disease, and areresponsible for osteolysis.

As used herein, “osteolysis” means refers to the degeneration and/ordissolution of bone. In some embodiments, osteolysis is the process ofbone resorption, whereby the bone salts can be withdrawn by a humoralmechanism and returned to the tissue fluids, leaving behind adecalcified bone matrix. Osteolysis can be caused, inter alia, bydisease, infection or ischemia.

Ossification (or, bone formation/mineralization) is the process by whichosteoblasts lay down bone. There are two processes resulting in theformation of normal, healthy bone tissue: intramembranous ossificationis the direct laying down of bone into the primitive connective tissue(mesenchyme), while endochondral ossification involves cartilage as aprecursor.

Osteoblasts are fibroblasts that express bone sialoprotein andosteocalcin. Osteoblasts produce a matrix of osteoid, which is composedmainly of Type I collagen. Osteoblasts are also responsible formineralization of this matrix. Zinc, copper and sodium are some of theminerals required in this process. The number of osteoblasts tends todecrease with age, affecting the balance of formation and resorption inthe bone tissue, and potentially leading to osteoporosis.

Disclosed herein, in certain embodiments, are methods of inhibiting boneresorption in an individual in need thereof, comprising administering tothe individual a composition comprising fetal support tissue or anextract thereof. In some embodiments, the bone resorption is undesiredor excessive. In some embodiments, inhibiting bone resorption promotesmineralization by osteoblasts. In some embodiments, inhibiting boneresorption inhibits undesired or excessive bone remodeling. In someembodiments, inhibiting bone resorption balances bone resorption andbone formation. Further disclosed herein, in certain embodiments, aremethods of treating a disease, disorder, or condition characterized byexcessive or undesired bone resorption, the method comprisingadministering to an individual in need thereof a composition comprisingsubstantially isolated amniotic membrane or an extract thereof.Additionally disclosed herein, in certain embodiments, are methods ofinhibiting osteoclast differentiation in an individual in need thereof,comprising administering to the individual a composition comprisingfetal support tissue or an extract thereof. In some embodiments, theosteoclast differentiation is undesired or excessive. In someembodiments, inhibiting osteoclast differentiation promotesdifferentiation of osteoblasts and mineralization by osteoblasts. Insome embodiments, inhibiting osteoclast differentiation inhibitsundesired or excessive bone remodeling. In some embodiments, inhibitingosteoclast differentiation balances bone resorption and bone formation.Also disclosed herein, in certain embodiments, are methods of treating adisease, disorder, or condition characterized by excessive or undesiredosteoclast differentiation, the method comprising administering to anindividual in need thereof a composition comprising substantiallyisolated amniotic membrane or an extract thereof.

In some embodiments, the methods comprise administering to an individualin need thereof a therapeutically-effective amount of a compositioncomprising a fetal support tissue product or an extract thereof.

In some embodiments, the fetal support tissue product is substantiallyisolated amnion. In some embodiments, the fetal support tissue productis substantially isolated PAM. In some embodiments, the fetal supporttissue product is substantially isolated UCAM. In some embodiments, thefetal support tissue product is substantially isolated placenta. In someembodiments, the fetal support tissue product is substantially isolatedumbilical cord. In some embodiments, the fetal support tissue product issubstantially isolated chorion or amnion-chorion. In some embodiments,the fetal support tissue product is frozen or previously frozen PAM. Insome embodiments, the fetal support tissue product is frozen orpreviously frozen UCAM. In some embodiments, the fetal support tissueproduct is frozen or previously frozen placenta. In some embodiments,the fetal support tissue product is frozen or previously frozenumbilical cord. In some embodiments, the fetal support tissue product isfrozen or previously frozen chorion or amnion-chorion. In someembodiments, the fetal support tissue product does not comprise a veinor an artery, a cell with metabolic activity, active HIV-1, activeHIV-2, active HTLV-1, active hepatitis B, active hepatitis C, activeWest Nile Virus, active cytomegalovirus, active human transmissiblespongiform encephalopathy, or active treponema pallidum. In someembodiments, the fetal support tissue product is obtained from a human,non-primate human, cow or pig. In some embodiments, the fetal supporttissue product is cryopreserved, lyophilized, terminally sterilized, ora combination thereof. In some embodiments, the fetal support tissueproduct is in the form of a pulverized powder or a homogenate.

In some embodiments, the methods comprise administering to an individualin need thereof a therapeutically-effective amount of a compositioncomprising substantially isolated HC-HA complex. In some embodiments,the substantially isolated HC-HA complex is derived from amnion (PAM orUCAM), placenta, umbilical cord, or chorion. In some embodiments, thesubstantially isolated HC-HA complex is derived from frozen orpreviously frozen placental amniotic membrane (PAM), frozen orpreviously frozen umbilical cord amniotic membrane (UCAM), frozen orpreviously frozen placenta, frozen or previously frozen umbilical cord,frozen or previously frozen chorion, frozen or previously frozenamnion-chorion, or any combinations thereof. In some embodiments, theHC-HA complex is HC-HAHC-HA obtained by a process comprising: (a)providing a reaction mixture comprising: (i) HA (e.g., HMW HA); (ii)IαI, wherein the IαI is optionally in serum or isolated from serum;(iii) TSG-6, wherein the TSG-6 is optionally recombinant; and (iv) PTX3,wherein the PTX3 is optionally recombinant; wherein at least one of HA,IαI, TSG-6, or PTX3 is optionally generated by a plurality of cellspresent in the reaction mixture; (b) incubating the reaction mixture fora period of time sufficient to produce HC-HA complex; and (c) isolatingand purifying the HC-HA complex.

In some embodiments, the composition further comprises apharmaceutically-acceptable diluents, excipient or carrier. In someembodiments, the composition is formulated as a solution, suspension,paste, ointment, oil, emulsion, microemulsion, cream, lotion, gel, orcombination thereof. In some embodiments, the composition comprisessubstantially-isolated HC-HA formulated into microspheres,microparticles, or liposomes. In some embodiments, the composition isformulated into PLGA copolymers. In some embodiments, the composition isformulated for controlled, sustained, or delayed release. In someembodiments, the composition is injected into an osteolytic joint. Insome embodiments, the composition is administered by a patch. In someembodiments, the patch is placed on an osteolytic bone or an osteolyticjoint. In some embodiments, the composition is administered by animplant. In some embodiments, the implant is placed on an osteolyticbone or an osteolytic joint. In some embodiments, the composition isformulated into an orthopaedic prosthesis.

Arthritis

Arthritis is a joint disorder that involves inflammation of one or morejoints. There are over 100 different forms of arthritis. The most commonform, osteoarthritis (degenerative joint disease), is a result of traumato the joint, infection of the joint, or age. Other arthritis forms arerheumatoid arthritis, psoriatic arthritis, and related autoimmunediseases. Septic arthritis is caused by joint infection. Symptoms commonto all arthritic disorders include pain, swelling, and joint stiffness,

Osteoarthritis is the most common form of arthritis. It affects both thelarger and the smaller joints of the body, for example the hands, feet,back, hip or knee. Osteoarthritis develops as a result of wear on jointsor injury to joints. Osteoarthritis begins in the cartilage andeventually causes the two opposing bones to erode into each other.Osteoarthritis typically affects the weight bearing joints such as theback, spine, and pelvis. Osteoarthritis is most commonly a disease ofthe elderly. More than 30 percent of females have some degree ofosteoarthritis by age 65. Risk factors for osteoarthritis include: priorjoint trauma, obesity, and a sedentary lifestyle.

Rheumatoid arthritis is a disorder in which the body's immune systemstarts to attack body tissues. In rheumatoid arthritis, most damageoccurs to the joint lining and cartilage which eventually results inerosion of two opposing bones. Rheumatoid arthritis often affects jointsin the fingers, wrists, knees and elbows. The disease is symmetrical(appears on both sides of the body) and can lead to severe deformity.Rheumatoid arthritis occurs mostly in people aged 20 and above.

Osteoclasts are often found in the affected joints of individuals witharthritis and exacerbate bone and joint destruction. Thus, disclosedherein, in certain embodiments, are methods of treating arthritis. Insome embodiments, the methods comprise administering to an individual inneed thereof a therapeutically-effective amount of a compositioncomprising a fetal support tissue product or an extract thereof. In someembodiments, the methods comprise administering to an individual in needthereof a therapeutically-effective amount of a composition comprisingsubstantially isolated HC-HA complex. In some embodiments, the arthritisis osteoarthritis, rheumatoid arthritis, psoriatic arthritis, or anycombination thereof.

In some embodiments, the methods comprise administering to an individualin need thereof a therapeutically-effective amount of a compositioncomprising a fetal support tissue product or an extract thereof.

In some embodiments, the fetal support tissue product is substantiallyisolated amnion. In some embodiments, the fetal support tissue productis substantially isolated PAM. In some embodiments, the fetal supporttissue product is substantially isolated UCAM. In some embodiments, thefetal support tissue product is substantially isolated placenta. In someembodiments, the fetal support tissue product is substantially isolatedumbilical cord. In some embodiments, the fetal support tissue product issubstantially isolated chorion or amnion-chorion. In some embodiments,the fetal support tissue product is frozen or previously frozen PAM. Insome embodiments, the fetal support tissue product is frozen orpreviously frozen UCAM. In some embodiments, the fetal support tissueproduct is frozen or previously frozen placenta. In some embodiments,the fetal support tissue product is frozen or previously frozenumbilical cord. In some embodiments, the fetal support tissue product isfrozen or previously frozen chorion or amnion-chorion. In someembodiments, the fetal support tissue product does not comprise a veinor an artery, a cell with metabolic activity, active HIV-1, activeHIV-2, active HTLV-1, active hepatitis B, active hepatitis C, activeWest Nile Virus, active cytomegalovirus, active human transmissiblespongiform encephalopathy, or active treponema pallidum. In someembodiments, the fetal support tissue product is obtained from a human,non-primate human, cow or pig. In some embodiments, the fetal supporttissue product is cryopreserved, lyophilized, terminally sterilized, ora combination thereof. In some embodiments, the fetal support tissueproduct is in the form of a pulverized powder or a homogenate.

In some embodiments, the methods comprise administering to an individualin need thereof a therapeutically-effective amount of a compositioncomprising substantially isolated HC-HA complex. In some embodiments,the substantially isolated HC-HA complex is isolated from amnion (PAM orUCAM), placenta, umbilical cord, or chorion. In some embodiments, thesubstantially isolated HC-HA complex is isolated from frozen orpreviously frozen placental amniotic membrane (PAM), frozen orpreviously frozen umbilical cord amniotic membrane (UCAM), frozen orpreviously frozen placenta, frozen or previously frozen umbilical cord,frozen or previously frozen chorion, frozen or previously frozenamnion-chorion, or any combinations thereof.

In some embodiments, the HC-HA complex is obtained by a processcomprising: (a) providing a reaction mixture comprising: (i) HA (e.g.,HMW HA); (ii) IαI, wherein the IαI is optionally in serum or isolatedfrom serum; (iii) TSG-6, wherein the TSG-6 is optionally recombinant;and (iv) PTX3, wherein the PTX3 is optionally recombinant; wherein atleast one of HA, IαI, TSG-6, or PTX3 is optionally generated by aplurality of cells present in the reaction mixture; (b) incubating thereaction mixture for a period of time sufficient to produce HC-HAcomplex; and (c) isolating and purifying the HC-HA complex.

In some embodiments, the composition further comprises apharmaceutically-acceptable diluents, excipient or carrier. In someembodiments, the composition is formulated as a solution, suspension,paste, ointment, oil, emulsion, microemulsion, cream, lotion, gel, orcombination thereof. In some embodiments, the composition comprisessubstantially-isolated HC-HA formulated into microspheres,microparticles, or liposomes. In some embodiments, the composition isformulated into PLGA copolymers. In some embodiments, the composition isformulated for controlled, sustained, or delayed release. In someembodiments, the composition is injected into an osteolytic joint. Insome embodiments, the composition is administered by a patch. In someembodiments, the patch is placed on an osteolytic bone or an osteolyticjoint. In some embodiments, the composition is administered by animplant. In some embodiments, the implant is placed on an osteolyticbone or an osteolytic joint. In some embodiments, the composition isformulated into an orthopaedic prosthesis.

In some embodiments, the methods further comprise administering to theindividual a calcium supplement. In some embodiments, the methodsfurther comprise administering to the individual an NSAID, acorticosteroid, hyaluronan injections, a DMARD, an analgesic, or anycombination thereof.

Osteoporosis

Osteoporosis is a bone disease characterized by reduction of bonemineral density (BMD), detioration of bone microarchitecture, andalteration of the amount and variety of proteins in bone. Osteoporosisis defined as a bone mineral density that is 2.5 standard deviations ormore below the mean peak bone mass (average of young, healthy adults) asmeasured by DXA. Osteoporosis is classified as primary type 1, primarytype 2, or secondary. The form of osteoporosis most common in womenafter menopause is referred to as primary type 1 or postmenopausalosteoporosis. Primary type 2 osteoporosis or senile osteoporosis occursafter age 75 and is seen in both females and males at a ratio of 2:1.Finally, secondary osteoporosis may arise at any age and affect men andwomen equally. This form of osteoporosis results from chronicpredisposing medical problems or disease, or prolonged use ofmedications such as glucocorticoids, when the disease is called steroid-or glucocorticoid-induced osteoporosis (SIOP or GIOP).

Osteoporosis results from excessive bone resorption by osteoclasts andinsufficient bone formation by osteoblasts. Thus, disclosed herein, incertain embodiments, are methods of treating osteoporosis. In someembodiments, the methods comprising administering to an individual inneed thereof a therapeutically-effective amount of a compositioncomprising a fetal support tissue product or an extract thereof. In someembodiments, the methods comprise administering to an individual in needthereof a therapeutically-effective amount of a composition comprisingsubstantially isolated HC-HA complex.

In some embodiments, the methods comprise administering to an individualin need thereof a therapeutically-effective amount of a compositioncomprising a fetal support tissue product or an extract thereof.

In some embodiments, the fetal support tissue product is substantiallyisolated amnion. In some embodiments, the fetal support tissue productis substantially isolated PAM. In some embodiments, the fetal supporttissue product is substantially isolated UCAM. In some embodiments, thefetal support tissue product is substantially isolated placenta. In someembodiments, the fetal support tissue product is substantially isolatedumbilical cord. In some embodiments, the fetal support tissue product issubstantially isolated chorion or amnion-chorion. In some embodiments,the fetal support tissue product is frozen or previously frozen PAM. Insome embodiments, the fetal support tissue product is frozen orpreviously frozen UCAM. In some embodiments, the fetal support tissueproduct is frozen or previously frozen placenta. In some embodiments,the fetal support tissue product is frozen or previously frozenumbilical cord. In some embodiments, the fetal support tissue product isfrozen or previously frozen chorion or amnion-chorion. In someembodiments, the fetal support tissue product does not comprise a veinor an artery, a cell with metabolic activity, active HIV-1, activeHIV-2, active HTLV-1, active hepatitis B, active hepatitis C, activeWest Nile Virus, active cytomegalovirus, active human transmissiblespongiform encephalopathy, or active treponema pallidum. In someembodiments, the fetal support tissue product is obtained from a human,non-primate human, cow or pig. In some embodiments, the fetal supporttissue product is cryopreserved, lyophilized, terminally sterilized, ora combination thereof. In some embodiments, the fetal support tissueproduct is in the form of a pulverized powder or a homogenate.

In some embodiments, the methods comprise administering to an individualin need thereof a therapeutically-effective amount of a compositioncomprising substantially isolated HC-HA complex. In some embodiments,the substantially isolated HC-HA complex is isolated from amnion (PAM orUCAM), placenta, umbilical cord, or chorion. In some embodiments, thesubstantially isolated HC-HA complex is isolated from frozen orpreviously frozen placental amniotic membrane (PAM), frozen orpreviously frozen umbilical cord amniotic membrane (UCAM), frozen orpreviously frozen placenta, frozen or previously frozen umbilical cord,frozen or previously frozen chorion, frozen or previously frozenamnion-chorion, or any combinations thereof.

In some embodiments, the HC-HA complex is obtained by a processcomprising: (a) providing a reaction mixture comprising: (i) HA (e.g.,HMW HA); (ii) IαI, wherein the IαI is optionally in serum or isolatedfrom serum; (iii) TSG-6, wherein the TSG-6 is optionally recombinant;and (iv) PTX3, wherein the PTX3 is optionally recombinant; wherein atleast one of HA, IαI, TSG-6, or PTX3 is optionally generated by aplurality of cells present in the reaction mixture; (b) incubating thereaction mixture for a period of time sufficient to produce HC-HAcomplex; and (c) isolating and purifying the HC-HA complex.

In some embodiments, the composition further comprises apharmaceutically-acceptable diluents, excipient or carrier. In someembodiments, the composition is formulated as a solution, suspension,paste, ointment, oil, emulsion, microemulsion, cream, lotion, gel, orcombination thereof. In some embodiments, the composition comprisessubstantially-isolated tissue product or HC-HA formulated intomicrospheres, microparticles, or liposomes. In some embodiments, thecomposition is formulated into PLGA copolymers. In some embodiments, thecomposition is formulated for controlled, sustained, or delayed release.In some embodiments, the composition is injected into an osteolyticjoint. In some embodiments, the composition is administered by a patch.In some embodiments, the patch is placed on an osteolytic bone or anosteolytic joint. In some embodiments, the composition is administeredby an implant. In some embodiments, the implant is placed on anosteolytic bone or an osteolytic joint. In some embodiments, thecomposition is formulated into an orthopaedic prosthesis. In someembodiments, the fetal support tissue product is in the form of asubstantially-flattened sheet. In some embodiments, the compositionfurther comprises a backing. In some embodiments, the composition iswrapped around an osteolytic bone or an osteolytic joint. In someembodiments, the methods further comprise administering to theindividual a calcium supplement. In some embodiments, the methodsfurther comprise administering to the individual a bisphosphonate, anestrogen analog, Raloxifene, Calcitonin, Teriparatide, calcium salts,sodium fluoride, RANKL inhibitors, Strontium ranelate, or anycombination thereof.

Alveolar Bone Degradation

Degradation of the alveolar bone around the teeth often results fromperiodontitis. If left untreated, alveolar bone degradation may resultin loss of teeth. A diagnosis of alveolar bone degradation isestablished by inspecting the soft gum tissues around the teeth with aprobe and evaluating x-ray films to determine the amount of bone lossaround the teeth.

Degradation of the alveolar bone is often exacerbated by excess boneresorption by osteoclasts and insufficient bone formation byosteoblasts. Thus, disclosed herein, in certain embodiments, are methodsof treating alveolar bone degradation. In some embodiments, the methodscomprising administering to an individual in need thereof atherapeutically-effective amount of a composition comprising a fetalsupport tissue product or an extract thereof. In some embodiments, themethods comprise administering to an individual in need thereof atherapeutically-effective amount of a composition comprisingsubstantially isolated HC-HA complex.

In some embodiments, the methods comprise administering to an individualin need thereof a therapeutically-effective amount of a compositioncomprising a fetal support tissue product or an extract thereof.

In some embodiments, the fetal support tissue product is substantiallyisolated amnion. In some embodiments, the fetal support tissue productis substantially isolated PAM. In some embodiments, the fetal supporttissue product is substantially isolated UCAM. In some embodiments, thefetal support tissue product is substantially isolated placenta. In someembodiments, the fetal support tissue product is substantially isolatedumbilical cord. In some embodiments, the fetal support tissue product issubstantially isolated chorion or amnion-chorion. In some embodiments,the fetal support tissue product is frozen or previously frozen PAM. Insome embodiments, the fetal support tissue product is frozen orpreviously frozen UCAM. In some embodiments, the fetal support tissueproduct is frozen or previously frozen placenta. In some embodiments,the fetal support tissue product is frozen or previously frozenumbilical cord. In some embodiments, the fetal support tissue product isfrozen or previously frozen chorion or amnion-chorion. In someembodiments, the fetal support tissue product does not comprise a veinor an artery, a cell with metabolic activity, active HIV-1, activeHIV-2, active HTLV-1, active hepatitis B, active hepatitis C, activeWest Nile Virus, active cytomegalovirus, active human transmissiblespongiform encephalopathy, or active treponema pallidum. In someembodiments, the fetal support tissue product is obtained from a human,non-primate human, cow or pig. In some embodiments, the fetal supporttissue product is cryopreserved, lyophilized, terminally sterilized, ora combination thereof. In some embodiments, the fetal support tissueproduct is in the form of a pulverized powder or a homogenate.

In some embodiments, the methods comprise administering to an individualin need thereof a therapeutically-effective amount of a compositioncomprising substantially isolated HC-HA complex. In some embodiments,the substantially isolated HC-HA complex is isolated from amnion (PAM orUCAM), placenta, umbilical cord, or chorion. In some embodiments, thesubstantially isolated HC-HA complex is isolated from frozen orpreviously frozen placental amniotic membrane (PAM), frozen orpreviously frozen umbilical cord amniotic membrane (UCAM), frozen orpreviously frozen placenta, frozen or previously frozen umbilical cord,frozen or previously frozen chorion, frozen or previously frozenamnion-chorion, or any combinations thereof.

In some embodiments, the HC-HA complex is obtained by a processcomprising: (a) providing a reaction mixture comprising: (i) HA (e.g.,HMW HA); (ii) IαI, wherein the IαI is optionally in serum or isolatedfrom serum; (iii) TSG-6, wherein the TSG-6 is optionally recombinant;and (iv) PTX3, wherein the PTX3 is optionally recombinant; wherein atleast one of HA, IαI, TSG-6, or PTX3 is optionally generated by aplurality of cells present in the reaction mixture; (b) incubating thereaction mixture for a period of time sufficient to produce HC-HAcomplex; and (c) isolating and purifying the HC-HA complex.

In some embodiments, the composition further comprises apharmaceutically-acceptable diluents, excipient or carrier. In someembodiments, the composition is formulated as a solution, suspension,paste, ointment, oil, emulsion, microemulsion, cream, lotion, gel, orcombination thereof. In some embodiments, the composition comprisessubstantially-isolated tissue product or HC-HA formulated intomicrospheres, microparticles, or liposomes. In some embodiments, thecomposition is formulated into PLGA copolymers. In some embodiments, thecomposition is formulated for controlled, sustained, or delayed release.In some embodiments, the composition is applied to the alveolar bone. Insome embodiments, the composition is administered by a patch. In someembodiments, the patch is placed on the alveolar bone. In someembodiments, the composition is administered by an implant. In someembodiments, the implant is placed on the alveolar bone. In someembodiments, the fetal support tissue product is in the form of asubstantially-flattened sheet. In some embodiments, the compositionfurther comprises a backing. In some embodiments, the composition iswrapped around the alveolar bone. In some embodiments, the methodsfurther comprise administering to the individual a calcium supplement.

Paget's Disease

Paget's disease is a chronic bone disorder resulting is enlarged andmisshapen bones. Bones in individual with Paget's Disease are oftenweakened due to an imbalance in bone resorption and bone formation,resulting in pain, misshapen bones, fractures, and arthritis in thejoints near the affected bones. Paget's disease typically is localizedto one or a few bones. Paget's Disease may result from a viral infectionor genetic factors.

Paget's disease proceeds in three stages: osteoclastic activity, mixedosteoclastic-osteoblastic activity, and exhaustive stage. The firststage is characterized by an increase in the rate of bone resorption atlocalized areas. These localized areas of osteolysis are seenradiologically as an advancing lytic wedge in long bones or osteoporosiscircumscripta in the skull. The osteolysis is followed by a compensatoryincrease in bone formation induced by osteoblasts recruited to the area.Bone is laid down in a disorganized and chaotic fashion rather than thenormal linear lamellar pattern. The resorbed bone is replaced and themarrow spaces are filled by an excess of fibrous connective tissue witha marked increase in blood vessels, causing the bone to becomehypervascular. The bone hypercellularity may then diminish, leaving adense “pagetic bone,” also known as burned-out Paget's disease.

Paget's disease results from excessive bone resorption and insufficientbone formation. Thus, disclosed herein, in certain embodiments, aremethods of treating Paget's Disease. In some embodiments, the methodscomprising administering to an individual in need thereof atherapeutically-effective amount of a composition comprising a fetalsupport tissue product or an extract thereof. In some embodiments, themethods comprise administering to an individual in need thereof atherapeutically-effective amount of a composition comprisingsubstantially isolated HC-HA complex.

In some embodiments, the methods comprise administering to an individualin need thereof a therapeutically-effective amount of a compositioncomprising a fetal support tissue product or an extract thereof.

In some embodiments, the fetal support tissue product is substantiallyisolated amnion. In some embodiments, the fetal support tissue productis substantially isolated PAM. In some embodiments, the fetal supporttissue product is substantially isolated UCAM. In some embodiments, thefetal support tissue product is substantially isolated placenta. In someembodiments, the fetal support tissue product is substantially isolatedumbilical cord. In some embodiments, the fetal support tissue product issubstantially isolated chorion or amnion-chorion. In some embodiments,the fetal support tissue product is frozen or previously frozen PAM. Insome embodiments, the fetal support tissue product is frozen orpreviously frozen UCAM. In some embodiments, the fetal support tissueproduct is frozen or previously frozen placenta. In some embodiments,the fetal support tissue product is frozen or previously frozenumbilical cord. In some embodiments, the fetal support tissue product isfrozen or previously frozen chorion or amnion-chorion. In someembodiments, the fetal support tissue product does not comprise a veinor an artery, a cell with metabolic activity, active HIV-1, activeHIV-2, active HTLV-1, active hepatitis B, active hepatitis C, activeWest Nile Virus, active cytomegalovirus, active human transmissiblespongiform encephalopathy, or active treponema pallidum. In someembodiments, the fetal support tissue product is obtained from a human,non-primate human, cow or pig. In some embodiments, the fetal supporttissue product is cryopreserved, lyophilized, terminally sterilized, ora combination thereof. In some embodiments, the fetal support tissueproduct is in the form of a pulverized powder or a homogenate.

In some embodiments, the methods comprise administering to an individualin need thereof a therapeutically-effective amount of a compositioncomprising substantially isolated HC-HA complex. In some embodiments,the substantially isolated HC-HA complex is isolated from amnion (PAM orUCAM), placenta, umbilical cord, or chorion. In some embodiments, thesubstantially isolated HC-HA complex is isolated from frozen orpreviously frozen placental amniotic membrane (PAM), frozen orpreviously frozen umbilical cord amniotic membrane (UCAM), frozen orpreviously frozen placenta, frozen or previously frozen umbilical cord,frozen or previously frozen chorion, frozen or previously frozenamnion-chorion, or any combinations thereof.

In some embodiments, the HC-HA complex is obtained by a processcomprising: (a) providing a reaction mixture comprising: (i) HA (e.g.,HMW HA); (ii) IαI, wherein the IαI is optionally in serum or isolatedfrom serum; (iii) TSG-6, wherein the TSG-6 is optionally recombinant;and (iv) PTX3, wherein the PTX3 is optionally recombinant; wherein atleast one of HA, IαI, TSG-6, or PTX3 is optionally generated by aplurality of cells present in the reaction mixture; (b) incubating thereaction mixture for a period of time sufficient to produce HC-HAcomplex; and (c) isolating and purifying the HC-HA complex.

In some embodiments, the composition further comprises apharmaceutically-acceptable diluents, excipient or carrier. In someembodiments, the composition is formulated as a solution, suspension,paste, ointment, oil, emulsion, microemulsion, cream, lotion, gel, orcombination thereof. In some embodiments, the composition comprisessubstantially-isolated tissue product or HC-HA formulated intomicrospheres, microparticles, or liposomes. In some embodiments, thecomposition is formulated into PLGA copolymers. In some embodiments, thecomposition is formulated for controlled, sustained, or delayed release.In some embodiments, the composition is injected into an osteolyticjoint. In some embodiments, the composition is administered by a patch.In some embodiments, the patch is placed on an osteolytic bone or anosteolytic joint. In some embodiments, the composition is administeredby an implant. In some embodiments, the implant is placed on anosteolytic bone or an osteolytic joint. In some embodiments, thecomposition is formulated into an orthopaedic prosthesis. In someembodiments, the fetal support tissue product is in the form of asubstantially-flattened sheet. In some embodiments, the compositionfurther comprises a backing. In some embodiments, the composition iswrapped around an osteolytic bone or an osteolytic joint. In someembodiments, the methods further comprise administering to theindividual a calcium supplement. In some embodiments, the methodsfurther comprise administering to the individual a bisphosphonate orcalcitonin.

Bone Tumors

As used herein, “bone tumor” means the uncontrolled and/or undesiredgrowth of tissue in bone. Bone tumors are either benign or malignant.They are further classified as “primary tumors”, which originate in boneor from bone-derived cells and tissues, and “secondary tumors” whichoriginate in other sites and metastasize to the bone.

Primary bone tumors may be neoplastic, developmental, traumatic,infectious, or inflammatory. Examples of benign bone tumors includeosteoma, osteoid osteoma, osteochondroma, osteoblastoma, enchondroma,giant cell tumor of bone, aneurysmal bone cyst, and fibrous dysplasia ofbone. Malignant primary bone tumors include osteosarcoma,chondrosarcoma, Ewing's sarcoma, fibrosarcoma, and other types.

Bone tumors often result in undesired bone degradation. Undesired bonedegradation may be caused by the presence of metastatic tumor cells thatstimulate formation and activation of osteoclasts. Undesired bonedegradation may also arise from the formation of osteoblasticmetastases. Thus, disclosed herein, in certain embodiments, are methodsof treating bone tumors. In some embodiments, the methods comprisingadministering to an individual in need thereof atherapeutically-effective amount of a composition comprising a fetalsupport tissue product or an extract thereof. In some embodiments, themethods comprise administering to an individual in need thereof atherapeutically-effective amount of a composition comprisingsubstantially isolated HC-HA complex.

In some embodiments, the methods comprise administering to an individualin need thereof a therapeutically-effective amount of a compositioncomprising a fetal support tissue product or an extract thereof.

In some embodiments, the fetal support tissue product is substantiallyisolated amnion. In some embodiments, the fetal support tissue productis substantially isolated PAM. In some embodiments, the fetal supporttissue product is substantially isolated UCAM. In some embodiments, thefetal support tissue product is substantially isolated placenta. In someembodiments, the fetal support tissue product is substantially isolatedumbilical cord. In some embodiments, the fetal support tissue product issubstantially isolated chorion or amnion-chorion. In some embodiments,the fetal support tissue product is frozen or previously frozen PAM. Insome embodiments, the fetal support tissue product is frozen orpreviously frozen UCAM. In some embodiments, the fetal support tissueproduct is frozen or previously frozen placenta. In some embodiments,the fetal support tissue product is frozen or previously frozenumbilical cord. In some embodiments, the fetal support tissue product isfrozen or previously frozen chorion or amnion-chorion. In someembodiments, the fetal support tissue product does not comprise a veinor an artery, a cell with metabolic activity, active HIV-1, activeHIV-2, active HTLV-1, active hepatitis B, active hepatitis C, activeWest Nile Virus, active cytomegalovirus, active human transmissiblespongiform encephalopathy, or active treponema pallidum. In someembodiments, the fetal support tissue product is obtained from a human,non-primate human, cow or pig. In some embodiments, the fetal supporttissue product is cryopreserved, lyophilized, terminally sterilized, ora combination thereof. In some embodiments, the fetal support tissueproduct is in the form of a pulverized powder or a homogenate.

In some embodiments, the methods comprise administering to an individualin need thereof a therapeutically-effective amount of a compositioncomprising substantially isolated HC-HA complex. In some embodiments,the substantially isolated HC-HA complex is isolated from amnion (PAM orUCAM), placenta, umbilical cord, or chorion. In some embodiments, thesubstantially isolated HC-HA complex is isolated from frozen orpreviously frozen placental amniotic membrane (PAM), frozen orpreviously frozen umbilical cord amniotic membrane (UCAM), frozen orpreviously frozen placenta, frozen or previously frozen umbilical cord,frozen or previously frozen chorion, frozen or previously frozenamnion-chorion, or any combinations thereof.

In some embodiments, the HC-HA complex is obtained by a processcomprising: (a) providing a reaction mixture comprising: (i) HA (e.g.,HMW HA); (ii) IαI, wherein the IαI is optionally in serum or isolatedfrom serum; (iii) TSG-6, wherein the TSG-6 is optionally recombinant;and (iv) PTX3, wherein the PTX3 is optionally recombinant; wherein atleast one of HA, IαI, TSG-6, or PTX3 is optionally generated by aplurality of cells present in the reaction mixture; (b) incubating thereaction mixture for a period of time sufficient to produce HC-HAcomplex; and (c) isolating and purifying the HC-HA complex.

In some embodiments, the composition further comprises apharmaceutically-acceptable diluents, excipient or carrier. In someembodiments, the composition is formulated as a solution, suspension,paste, ointment, oil, emulsion, microemulsion, cream, lotion, gel, orcombination thereof. In some embodiments, the composition comprisessubstantially-isolated tissue product or HC-HA formulated intomicrospheres, microparticles, or liposomes. In some embodiments, thecomposition is formulated into PLGA copolymers. In some embodiments, thecomposition is formulated for controlled, sustained, or delayed release.In some embodiments, the composition is injected into an osteolyticjoint. In some embodiments, the composition is administered by a patch.In some embodiments, the patch is placed on an osteolytic bone or anosteolytic joint. In some embodiments, the composition is administeredby an implant. In some embodiments, the implant is placed on anosteolytic bone or an osteolytic joint. In some embodiments, thecomposition is formulated into an orthopaedic prosthesis. In someembodiments, the fetal support tissue product is in the form of asubstantially-flattened sheet. In some embodiments, the compositionfurther comprises a backing. In some embodiments, the composition iswrapped around an osteolytic bone or an osteolytic joint. In someembodiments, the methods further comprise administering to theindividual a calcium supplement. In some embodiments, the methodsfurther comprise administering to the individual a chemotherapeuticagent, a bisphosphinate, Metastron, or any combination thereof.

Fetal Support Tissue Products

Disclosed herein, in certain embodiments, are methods of inhibitingosteoclast differentiation in an individual in need thereof, comprisingadministering to the individual a composition comprising product fetalsupport tissue product or an extract thereof. Disclosed herein, incertain embodiments, are methods of promoting mineralization byosteoblasts in an individual in need thereof, comprising administeringto the individual a composition comprising a fetal support tissueproduct or an extract thereof. In some embodiments, the fetal supporttissue product is substantially isolated placental amniotic membrane(PAM), umbilical cord amniotic membrane (UCAM), placenta, umbilicalcord, chorion, amnion-chorion, or any combinations thereof. Disclosedherein, in certain embodiments, are methods of inhibiting boneresorption in an individual in need thereof, comprising administering tothe individual a composition comprising a fetal support tissue productor an extract thereof. Disclosed herein, in certain embodiments, aremethods of inhibiting bone remodeling in an individual in need thereof,comprising administering to the individual a composition comprising afetal support tissue product or an extract thereof. Disclosed herein, incertain embodiments, are methods of balancing bone resorption and boneformation, comprising administering to an individual in need thereof atherapeutically-effective amount of a composition comprising a fetalsupport tissue product or an extract thereof. Disclosed herein, incertain embodiments, are methods of treating a disease, disorder, orcondition characterized by excessive or undesired osteoclastdifferentiation, the methods comprising administering to an individualin need thereof a therapeutically-effective amount of a compositioncomprising a fetal support tissue product or an extract thereof.Disclosed herein, in certain embodiments, are methods of treating adisease, disorder, or condition characterized by excessive or undesiredbone absorption by osteoclasts, the methods comprising administering toan individual in need thereof a therapeutically-effective amount of acomposition comprising a fetal support tissue product or an extractthereof. Disclosed herein, in certain embodiments, are methods oftreating a disease, disorder, or condition characterized by deficient ordefective bone formation, the methods comprising administering to anindividual in need thereof a therapeutically-effective amount of acomposition comprising a fetal support tissue product or an extractthereof. Disclosed herein, in certain embodiments, are methods oftreating arthritis, the methods comprising administering to anindividual in need thereof a therapeutically-effective amount of acomposition comprising a fetal support tissue product or an extractthereof. Disclosed herein, in certain embodiments, are methods oftreating osteoporosis, the methods comprising administering to anindividual in need thereof a therapeutically-effective amount of acomposition comprising a fetal support tissue product or an extractthereof. Disclosed herein, in certain embodiments, are methods oftreating alveolar bone degradation, the methods comprising administeringto an individual in need thereof a therapeutically-effective amount of acomposition comprising a fetal support tissue product or an extractthereof. Disclosed herein, in certain embodiments, are methods oftreating Paget's disease, the methods comprising administering to anindividual in need thereof a therapeutically-effective amount of acomposition comprising a fetal support tissue product or an extractthereof. Disclosed herein, in certain embodiments, are methods oftreating a bone tumor, the methods comprising administering to anindividual in need thereof a therapeutically-effective amount of acomposition comprising a fetal support tissue product or an extractthereof. In some embodiments of any of the aforementioned methods, thefetal support tissue product is substantially isolated amnion. In someembodiments of any of the aforementioned methods, the fetal supporttissue product is substantially isolated PAM. In some embodiments of anyof the aforementioned methods, the fetal support tissue product issubstantially isolated UCAM. In some embodiments of any of theaforementioned methods, the fetal support tissue product issubstantially isolated placenta. In some embodiments of any of theaforementioned methods, the fetal support tissue product issubstantially isolated umbilical cord. In some embodiments of any of theaforementioned methods, the fetal support tissue product issubstantially isolated chorion or amnion-chorion. In some embodiments ofany of the aforementioned methods, the fetal support tissue product isfrozen or previously frozen PAM. In some embodiments of any of theaforementioned methods, the fetal support tissue product is frozen orpreviously frozen UCAM. In some embodiments of any of the aforementionedmethods, the fetal support tissue product is frozen or previously frozenplacenta. In some embodiments of any of the aforementioned methods, thefetal support tissue product is frozen or previously frozen umbilicalcord. In some embodiments of any of the aforementioned methods, thefetal support tissue product is frozen or previously frozen chorion oramnion-chorion. In some embodiments of any of the aforementionedmethods, the fetal support tissue product is in the form of a sheet,pulverized powder or a homogenate.

Umbilical Cord and UCAM

Umbilical cord is recovered from any suitable source (e.g., a hospitalor tissue bank). Umbilical cord can be obtained from any mammal, such asa human, non-human primate, cow or pig.

The umbilical cord is kept frozen (e.g., at or below 0° C.) until donorand specimen eligibility has been determined. In some embodiments,freezing the umbilical cord kills substantially all cells found in theumbilical cord. In some embodiments, freezing the umbilical cord killssubstantially all cells found in the umbilical cord while maintaining orincreasing the biological activity of the umbilical cord relative tofresh (i.e., non-frozen) umbilical cord. In some embodiments, freezingthe umbilical cord results in the loss of metabolic activity insubstantially all cells found in the umbilical cord. In someembodiments, freezing the umbilical cord results in the loss ofmetabolic activity in substantially all cells found in the umbilicalcord while maintaining or increasing the biological activity of theumbilical cord (e.g., its anti-inflammatory, anti-scarring,anti-antigenic, and anti-adhesion properties) relative to fresh (i.e.,non-frozen) umbilical cord. In some embodiments, the umbilical cord isnot frozen. If the umbilical cord is not frozen, it is processed asdescribed below immediately. In some embodiments, the umbilical cord isdried (e.g., by lyophilization). In some embodiments, drying theumbilical cord kills substantially all cells found in the umbilicalcord. In some embodiments, drying the umbilical cord results in the lossof metabolic activity in substantially all cells found in the umbilicalcord.

All processing is done following Good Tissue Practices (GTP) to ensurethat no contaminants are introduced into the UCAM products.

The umbilical cord is tested for HIV-1, HIV-2, HTLV-1, hepatitis B andC, West Nile virus, cytomegalovirus, human transmissible spongiformencephalopathy (e.g., Creutzfeldt-Jakob disease) and treponema pallidumusing FDA licensed screening test. Any indication that the tissue iscontaminated with HIV-1, HIV-2, HTLV-1, hepatitis B and C, West Nilevirus, or cytomegalovirus results in the immediate quarantine andsubsequent destruction of the tissue specimen.

Further, the donor's medical records are examined for risk factors forand clinical evidence of hepatitis B, hepatitis C, or HIV infection. Anyindication that the donor has risk factors for, and/or clinical evidenceof, infection with HIV-1, HIV-2, HTLV-1, hepatitis B and C, West Nilevirus, cytomegalovirus, human transmissible spongiform encephalopathy(e.g., Creutzfeldt-Jakob disease) and treponema pallidum results in theimmediate quarantine and subsequent destruction of the tissue specimen.

In some embodiments, substantially all of the blood is removed from theumbilical cord. In some embodiments, substantially all of the blood isremoved from the umbilical cord before the umbilical cord is frozen. Insome embodiments, blood is not removed from the UC. In some embodiments,blood is not removed from the umbilical cord before the umbilical cordis frozen.

In some embodiments, the umbilical cord tissue is washed with bufferwith agitation to remove excess blood and tissue. In some embodiments,washing with agitation reduces the wash time. In some embodiments, theumbilical cord is contacted with an isotonic buffer. In someembodiments, the umbilical cord is contacted with saline, PBS, PBS 1X,Ringer's solution, Hartmann's solution, TRIS-buffered saline,HEPES-buffered saline, EBSS, HBSS, Tyrode's salt Solution, Gey'sBalanced Salt Solution, DMEM, EMEM, GMEM, RPMI, or any combinationsthereof.

In some embodiments, the umbilical cord is cut into multiple sheets. Thesize of the sheets depends on the desired use of the product derivedfrom the umbilical cord.

In some embodiments, the section of the umbilical cord is not cutfurther. In some embodiments, the section of the umbilical cord is cutinto smaller portions. Optionally, in some embodiments, additional cutsare made in the Wharton's Jelly to help shape the UC.

In some embodiments, the cut umbilical cord tissue is optionally washedagain with buffer to further remove excess blood and tissue.

In some embodiments, part or all of the Wharton's Jelly is removed fromthe umbilical cord. The desired thickness of the fetal support tissueproduct determines how much, if any, of the Wharton's Jelly is removed.In some embodiments, the Wharton's Jelly is not removed.

The umbilical cord comprises two arteries (the umbilical arteries) andone vein (the umbilical vein). In some embodiments, the vein andarteries are removed from the umbilical cord. In certain instances, thevein and arteries are surrounded (or suspended or buried) within theWharton's Jelly. In some embodiments, the vein and arteries are removedconcurrently with the removal of the Wharton's Jelly. In someembodiments, the veins and arteries are not removed.

If a UCAM sheet is the desired fetal support tissue product, thengeneration of the UCAM sheet comprises substantially isolating the UCAMfrom the umbilical cord. After substantially pure UCAM has beenobtained, the UCAM is optionally washed with buffer to remove excessblood and tissue.

In some embodiments, isolated UCAM is flattened following separationfrom the umbilical cord, generating a flat UCAM sheet comprisingisolated UCAM. In some embodiments, the isolated UCAM is rolledfollowing separation from the umbilical cord, generating a tubular UCAMsheet. In some embodiments, the UCAM sheets are in any suitable shape(e.g., a square, a circle, a triangle, a rectangle) and into anysuitable size.

In some embodiments, the UCAM sheets are contacted with a buffer toremove substantially all remaining red blood cells. In some embodiments,the chorion or amnion-chorion sheets are contacted with an isotonicbuffer. In some embodiments, the chorion or amnion-chorion sheets arecontacted with saline, PBS, PBS 1X, Ringer's solution, Hartmann'ssolution, TRIS-buffered saline, HEPES-buffered saline, EBSS, HBSS,Tyrode's salt Solution, Gey's Balanced Salt Solution, DMEM, EMEM, GMEM,RPMI, or any combinations thereof.

In some embodiments, multiple layers of isolated UCAM are combined togenerate a layered UCAM sheet. The layered UCAM sheet is any suitablethickness. In some embodiments, the layered UCAM sheet comprises two,three, four, five, six, seven, eight, nine, or ten layers of isolatedUCAM. In some embodiments, the layered UCAM sheet comprises more thanten layers of isolated UCAM.

In some embodiments, the UCAM sheet is optionally contacted with asubstrate (i.e., a supportive backing). In some embodiments, UCAM sheetis not contacted with a substrate. In some embodiments, the UCAM sheetdoes not require a particular orientation relative to the substrate(i.e., any side of the UCAM may be in contact with the substrate). Insome embodiments, the UCAM sheet is orientated such that the epitheliallayer is in contact with the substrate.

In some embodiments, the natural structural integrity of the umbilicalcord (or, any isolated parts thereof, e.g., UCAM) is maintained in thesheet. In some embodiments, the natural biological activity of theumbilical cord (or, any isolated parts thereof, e.g., UCAM) ismaintained in the sheet. In some embodiments, the natural structuralintegrity and the natural biological activity of the umbilical cord (or,any isolated parts thereof, e.g., UCAM) is maintained in the sheet.

In some embodiments, a powder or homogenate is made by: obtainingumbilical cord, processing the umbilical cord (see above for processingmethods), and grinding (or pulverizing) or homogenizing the umbilicalcord. In some embodiments, the powder or homogenate is made with wholeumbilical cord. Optionally, the umbilical cord is further processed byremoving Wharton's Jelly and/or the umbilical arteries and veins.Alternatively, the Wharton's Jelly and/or the umbilical arteries andveins are note removed. In some embodiments, the powder or homogenate ismade by isolating the UCAM from the rest of the umbilical cord. In someembodiments, the natural biological activity of the umbilical cord (or,any isolated parts thereof) is substantially maintained.

In some embodiments, the powder or homogenate is prepared by anysuitable method. In some embodiments, the powder or homogenate isprepared by use of a homogenizer (e.g., an ultrasonic homogenizer),sonicator, pulverizer, Warring blender, grinding mill/jar, bead beater,or any combination thereof.

In some embodiments, the powder or homogenate is prepared by use of agrinding jar. In some embodiments, the umbilical cord (or, any isolatedparts thereof) is lyophilized prior to being placed in the grinding jar.A grinding ball is dropped in the grinding jar and the grinding jar issealed. The grinding jar is immersed into liquid nitrogen for 5 min andthen placed in a mill and ground at a 30 Hz grinding cycle for 4 min.

In some embodiments, the umbilical cord (or, any isolated parts thereof)is lyophilized by any suitable method (e.g., exposure to a liquid gas,placement in a freezer). In some embodiments, the umbilical cord (or,any isolated parts thereof) is placed in the vacuum chamber of alyophilization device until all or substantially all fluid (e.g., water)has been removed. In some embodiments, the umbilical cord (or, anyisolated parts thereof) is lyophilized following freezing (e.g.,exposure to a temperature below 0° C., −20° C., −40° C., −50° C., −60°C., −70° C., −75° C., −80° C., −90° C., −100° C.).

In some embodiments, the natural biological activity of the umbilicalcord (or, any isolated parts thereof) is maintained in the powder orhomogenate.

In some embodiments, an extract is made from the umbilical cord or anyisolated parts thereof. In some embodiments, the natural biologicalactivity of the umbilical cord (or, any isolated parts thereof) ismaintained in the extract. In some embodiments, a homogenate or powderis made as described above. In some embodiments, the homogenate orpowder is centrifuged to generate an extract (i.e., a chorion extract oran amnion-chorion extract). Any suitable method of centrifugation may beused. In some embodiments, the extract comprises the supernatant. Insome embodiments, the extract comprises the precipitant. In someembodiments, the extract is subject to additional extraction methods(e.g., HABP affinity chromotography, or immuno affinity chromatography).

In some embodiments, the method of making the extract comprises: (a)mixing the homogenate or powder with cold PBS buffer without proteaseinhibitors, to generate a PBS mixture, (b) centrifuging the PBS mixture,and (c) isolating the extract, to generate an isolated extract. In someembodiments, the cold PBS buffer and tissue product are combined in a1:1 ratio. In some embodiments, the PBS mixture is centrifuged at48,000×g 4° C. for 30 min.

In some embodiments, the method of making the extract further comprisespurifying the extract. The number of purification steps depends on thedesired purity. In some embodiments, the purification comprises at least2 rounds of ultracentrifugation. In some embodiments, the purificationcomprises more 2 rounds of ultracentrifugation. In some embodiments, thepurification comprises at least 4 rounds of ultracentrifugation. In someembodiments, the method of purifying the isolated extract comprises: (d)dissolving the isolated extract in CsCl/4M guanidine HCl at the initialdensity of 1.35 g/ml, to generate a CsCl mixture, (e) centrifuging theCsCl mixture at 125,000×g for 48 h at 15° C., to generate a firstpurified extract, (f) extracting the first purified extract anddialyzing it against distilled water to remove CsCl and guanidine HCl,to generate a dialysate. In some embodiments, the method of purifyingthe isolated extract further comprises (g) mixing the dialysate with 3volumes of 95% (v/v) ethanol containing 1.3% (w/v) potassium acetate at0° C. for 1 h, to generate a first dialysate/ethanol mixture, (h)centrifuging the first dialysate/ethanol mixture at 15,000×g, togenerate a second purified extract, and (i) extracting the secondpurified extract. In some embodiments, the method of purifying theisolated extract further comprises: (j) washing the second purifiedextract with ethanol (e.g., 70% ethanol), to generate a second purifiedextract/ethanol mixture; (k) centrifuging the second purifiedextract/ethanol mixture, to generate a third purified extract; and (1)extracting the third purified extract. In some embodiments, the methodof purifying the isolated extract further comprises: (m) washing thethird purified extract with ethanol (e.g., 70% ethanol), to generate athird purified extract/ethanol mixture; (n) centrifuging the thirdpurified extract/ethanol mixture, to generate a forth purified extract;and (O) extracting the forth purified extract. In some embodiments, thepurified extract comprises HC-HA complex.

Placenta and Placental Amniotic Membrane (PAM)

Placenta is recovered from any suitable source (e.g., a hospital ortissue bank). Placenta can be obtained from any mammal, such as a human,non-human primate, cow or pig. The placenta may be frozen, previouslyfrozen, or fresh (i.e., not frozen).

Where the placenta is not processed into a pulverized immediately afterit has been obtained, it is processed for storage (e.g., it is frozen ordried). In some embodiments, the placenta is frozen for storage. In someembodiments, the placenta is frozen at or below 0° C. In someembodiments, the placenta is frozen until donor and specimen eligibilityhas been determined. In some embodiments, the placenta is placed in acryo-preservative before being frozen. In some embodiments, freezing theplacenta kills substantially all cells found in the placenta. In someembodiments, freezing the placenta kills substantially all cells foundin the placenta while maintaining or increasing the biological activityof the placenta relative to fresh (i.e., non-frozen) placenta. In someembodiments, freezing the placenta results in the loss of metabolicactivity in substantially all cells found in the placenta. In someembodiments, freezing the placenta results in the loss of metabolicactivity in substantially all cells found in the placenta whilemaintaining or increasing the biological activity of the placentarelative to fresh (i.e., non-frozen) placenta. If the placenta is notfrozen, it is processed as described below immediately. In someembodiments, the placenta is dried (e.g., by lyophilization). In someembodiments, drying the placenta kills substantially all cells found inthe placenta. In some embodiments, drying the placenta results in theloss of metabolic activity in substantially all cells found in theplacenta.

All processing is done following Good Tissue Practices (GTP) to ensurethat no contaminants are introduced into the tissue grafts or PAM.

The placenta is tested for HIV-1, HIV-2, HTLV-1, hepatitis B and C, WestNile virus, cytomegalovirus, human transmissible spongiformencephalopathy (e.g., Creutzfeldt-Jakob disease) and treponema pallidumusing FDA licensed screening test. Any indication that the tissue iscontaminated with HIV-1, HIV-2, HTLV-1, hepatitis B and C, West Nilevirus, or cytomegalovirus results in the immediate quarantine andsubsequent destruction of the tissue specimen.

Further, the donor's medical records are examined for risk factors forand clinical evidence of hepatitis B, hepatitis C, or HIV infection. Anyindication that the donor has risk factors for, and/or clinical evidenceof, infection with HIV-1, HIV-2, HTLV-1, hepatitis B and C, West Nilevirus, cytomegalovirus, human transmissible spongiform encephalopathy(e.g., Creutzfeldt-Jakob disease) and treponema pallidum results in theimmediate quarantine and subsequent destruction of the tissue specimen.

In some embodiments, the PAM is not isolated from the placenta beforefurther processing begins. In some embodiments, the PAM is isolated fromthe placenta (generating isolated PAM) before further processing begins,

In some embodiments, substantially all of the blood is removed from theplacenta. In some embodiments, some blood is removed from placenta. Insome embodiments, the blood is not removed from the placenta.

In some embodiments, the placenta is washed with buffer with agitationto remove excess blood and tissue. In some embodiments, washing withagitation reduces the wash time.

In some embodiments, the placenta is contacted with a buffer. In someembodiments, the placenta is contacted with an isotonic buffer. In someembodiments, the placenta is contacted with saline, PBS, PBS 1X,Ringer's solution, Hartmann's solution, TRIS-buffered saline,HEPES-buffered saline, EBSS, HBSS, Tyrode's salt Solution, Gey'sBalanced Salt Solution, DMEM, EMEM, GMEM, RPMI, or any combinationsthereof.

In some embodiments, the placenta is washed with an isotonic buffer ortissue culture media, and any suitable antibiotic. In some embodiments,the antibiotic is ciprofloxacin, amphotericin B, penicillin,streptomycin, neomycin or a combination thereof. In some embodiments,the antibiotic is ciprofloxacin and amphotericin B. In some embodiments,the antibiotic is penicillin, streptomycin, neomycin, and amphotericinB.

In some embodiments, the placenta is cut into multiple sections (e.g.,using a scalpel). The size of the sections depends on the desired use ofthe product derived from the placenta.

In some embodiments, the cut placenta tissue is optionally washed againwith buffer to further remove excess blood and tissue.

If a PAM sheet is the desired fetal support tissue product, thengeneration of the PAM sheet comprises any or all of the following stepsin addition to the preceding steps.

In some embodiments, PAM is substantially isolated from the placenta.After substantially isolated PAM has been obtained, the PAM isoptionally washed with buffer to remove excess blood and tissue.

In some embodiments, isolated PAM is cut into in any suitable shape(e.g., a square, a circle, a triangle, a rectangle) and into anysuitable sizes.

In some embodiments, the PAM sheets are contacted with a buffer toremove substantially all remaining red blood cells. In some embodiments,the chorion or amnion-chorion sheets are contacted with an isotonicbuffer. In some embodiments, the chorion or amnion-chorion sheets arecontacted with saline, PBS, PBS 1X, Ringer's solution, Hartmann'ssolution, TRIS-buffered saline, HEPES-buffered saline, EBSS, HBSS,Tyrode's salt Solution, Gey's Balanced Salt Solution, DMEM, EMEM, GMEM,RPMI, or any combinations thereof.

In some embodiments, multiple layers of isolated PAM are combined togenerate a layered PAM sheet. The layered PAM sheet is any suitablethickness. In some embodiments, the layered PAM sheet comprises two,three, four, five, six, seven, eight, nine, or ten layers of isolatedPAM. In some embodiments, the layered PAM sheet comprises more than tenlayers of isolated PAM.

In some embodiments, the PAM sheet is optionally contacted with asubstrate (i.e., a supportive backing). In some embodiments, PAM sheetis not contacted with a substrate. In some embodiments, the PAM sheetdoes not require a particular orientation relative to the substrate(i.e., any side of the PAM may be in contact with the substrate). Insome embodiments, the PAM sheet is orientated such that the epitheliallayer is in contact with the substrate.

In some embodiments, the natural structural integrity of the placenta(or any isolated parts thereof; e.g., PAM) is maintained in the sheet.In some embodiments, the natural biological activity of the placenta (orany isolated parts thereof) is maintained in the sheet. In someembodiments, the natural structural integrity and natural biologicalactivity of the placenta (or any isolated parts thereof; e.g., PAM) ismaintained in the sheet.

In some embodiments, a powder or homogenate is made by: obtainingplacenta, processing the placenta (see above for processing methods),and grinding (or pulverizing) or homogenizing the placenta. In someembodiments, the powder or homogenate is made with whole placenta. Insome embodiments, the powder or homogenate is made by isolating the PAMfrom the rest of the placenta. In some embodiments, the naturalbiological activity of the placenta (or, any isolated parts thereof) issubstantially maintained.

In some embodiments, the powder or homogenate is prepared by anysuitable method. In some embodiments, the powder or homogenate isprepared by use of a homogenizer (e.g., an ultrasonic homogenizer),sonicator, pulverizer, Warring blender, grinding mill/jar, bead beater,or any combination thereof.

In some embodiments, the powder or homogenate is prepared by use of agrinding jar. In some embodiments, the placenta (or, any isolated partsthereof) is lyophilized prior to being placed in the grinding jar. Agrinding ball is dropped in the grinding jar and the grinding jar issealed. The grinding jar is immersed into liquid nitrogen for 5 min andthen placed in a mill and ground at a 30 Hz grinding cycle for 4 min.

In some embodiments, the placenta (or, any isolated parts thereof) islyophilized by any suitable method (e.g., exposure to a liquid gas,placement in a freezer). In some embodiments, the placenta (or, anyisolated parts thereof) is placed in the vacuum chamber of alyophilization device until all or substantially all fluid (e.g., water)has been removed. In some embodiments, the placenta (or, any isolatedparts thereof) is lyophilized following freezing (e.g., exposure to atemperature below 0° C., −20° C., −40° C., −50° C., −60° C., −70° C.,−75° C., −80° C., −90° C., −100° C.).

In some embodiments, the natural biological activity of the placenta orany isolated parts thereof is maintained in a powder or homogenate.

In some embodiments, an extract is made from the placenta or anyisolated parts thereof. In some embodiments, the natural biologicalactivity of the placenta or any isolated parts thereof is maintained inan extract. In some embodiments, a homogenate or powder is made asdescribed above. In some embodiments, the homogenate or powder iscentrifuged to generate an extract (i.e., a chorion extract or anamnion-chorion extract). Any suitable method of centrifugation may beused. In some embodiments, the extract comprises the supernatant. Insome embodiments, the extract comprises the precipitant. In someembodiments, the extract is subject to additional extraction methods(e.g., HABP affinity chromotography, or immunoaffinity chromatography).

In some embodiments, the method of making the extract comprises: (a)mixing the homogenate or powder with cold PBS buffer without proteaseinhibitors, to generate a PBS mixture, (b) centrifuging the PBS mixture,and (c) isolating the extract, to generate an isolated extract. In someembodiments, the cold PBS buffer and tissue product are combined in a1:1 ratio. In some embodiments, the PBS mixture is centrifuged at48,000×g 4° C. for 30 min.

In some embodiments, the method of making the extract further comprisespurifying the extract. The number of purification steps depends on thedesired purity. In some embodiments, the purification comprises at least2 rounds of ultracentrifugation. In some embodiments, the purificationcomprises more 2 rounds of ultracentrifugation. In some embodiments, thepurification comprises at least 4 rounds of ultracentrifugation. In someembodiments, the method of purifying the isolated extract comprises: (d)dissolving the isolated extract in CsCl/4M guanidine HCl at the initialdensity of 1.35 g/ml, to generate a CsCl mixture, (e) centrifuging theCsCl mixture at 125,000×g for 48 h at 15° C., to generate a firstpurified extract, (f) extracting the first purified extract anddialyzing it against distilled water to remove CsCl and guanidine HCl,to generate a dialysate. In some embodiments, the method of purifyingthe isolated extract further comprises (g) mixing the dialysate with 3volumes of 95% (v/v) ethanol containing 1.3% (w/v) potassium acetate at0° C. for 1 h, to generate a first dialysate/ethanol mixture, (h)centrifuging the first dialysate/ethanol mixture at 15,000×g, togenerate a second purified extract, and (i) extracting the secondpurified extract. In some embodiments, the method of purifying theisolated extract further comprises: (j) washing the second purifiedextract with ethanol (e.g., 70% ethanol), to generate a second purifiedextract/ethanol mixture; (k) centrifuging the second purifiedextract/ethanol mixture, to generate a third purified extract; and (1)extracting the third purified extract. In some embodiments, the methodof purifying the isolated extract further comprises: (m) washing thethird purified extract with ethanol (e.g., 70% ethanol), to generate athird purified extract/ethanol mixture; (n) centrifuging the thirdpurified extract/ethanol mixture, to generate a forth purified extract;and (o) extracting the forth purified extract. In some embodiments, thepurified extract comprises HC-HA complex.

Chorion and Amnion-Chorion

Placenta comprising chorion is recovered from any suitable source (e.g.,a hospital or tissue bank). Placenta can be obtained from any mammal,such as a human, non-human primate, cow or pig. The placenta may befrozen, previously frozen, or fresh (i.e., not frozen).

In some embodiments, the chorion is immediately isolated from theplacenta. In some embodiments, the chorion is not immediately isolatedfrom the placenta. In some embodiments, the chorion or amnion-chorion(or, placenta comprising chorion) is frozen for storage. In someembodiments, the chorion or amnion-chorion (or, placenta comprisingchorion) is frozen at or below 0° C. In some embodiments, the chorion oramnion-chorion (or, placenta comprising chorion) is frozen until donorand specimen eligibility has been determined. In some embodiments, thechorion or amnion-chorion (or, placenta comprising chorion) is placed ina cryo-preservative before being frozen. In some embodiments, freezingthe chorion or amnion-chorion (or, placenta comprising chorion) killssubstantially all cells found in the chorion or amnion-chorion. In someembodiments, freezing the chorion or amnion-chorion (or, placentacomprising chorion) kills substantially all cells found in the chorionor amnion-chorion while maintaining or increasing the biologicalactivity of the chorion or amnion-chorion relative to fresh (i.e.,non-frozen) chorion or amnion-chorion. In some embodiments, freezing thechorion or amnion-chorion (or, placenta comprising chorion) results inthe loss of metabolic activity in substantially all cells found in thechorion or amnion-chorion. In some embodiments, freezing the chorion oramnion-chorion (or, placenta comprising chorion) results in the loss ofmetabolic activity in substantially all cells found in the chorion oramnion-chorion while maintaining or increasing the biological activityof the chorion or amnion-chorion relative to fresh (i.e., non-frozen)chorion or amnion-chorion. If the chorion or amnion-chorion is notfrozen, it is processed as described below immediately. In someembodiments, the chorion or amnion-chorion (or, placenta comprisingchorion) is dried (e.g., by lyophilization). In some embodiments, dryingthe chorion or amnion-chorion (or, placenta comprising chorion) killssubstantially all cells found in the chorion or amnion-chorion. In someembodiments, drying the chorion or amnion-chorion (or, placentacomprising chorion) results in the loss of metabolic activity insubstantially all cells found in the chorion or amnion-chorion.

All processing is done following Good Tissue Practices (GTP) to ensurethat no contaminants are introduced into the tissue grafts or PAM.

The chorion or amnion-chorion (or, placenta comprising chorion) istested for HIV-1, HIV-2, HTLV-1, hepatitis B and C, West Nile virus,cytomegalovirus, human transmissible spongiform encephalopathy (e.g.,Creutzfeldt-Jakob disease) and treponema pallidum using FDA licensedscreening test. Any indication that the tissue is contaminated withHIV-1, HIV-2, HTLV-1, hepatitis B and C, West Nile virus, orcytomegalovirus results in the immediate quarantine and subsequentdestruction of the tissue specimen.

Further, the donor's medical records are examined for risk factors forand clinical evidence of hepatitis B, hepatitis C, or HIV infection. Anyindication that the donor has risk factors for, and/or clinical evidenceof, infection with HIV-1, HIV-2, HTLV-1, hepatitis B and C, West Nilevirus, cytomegalovirus, human transmissible spongiform encephalopathy(e.g., Creutzfeldt-Jakob disease) and treponema pallidum results in theimmediate quarantine and subsequent destruction of the tissue specimen.

In some embodiments, the chorion or amnion-chorion is not isolated fromthe placenta before further processing begins. In some embodiments, thechorion or amnion-chorion is isolated from the placenta before furtherprocessing begins.

In some embodiments, substantially all of the blood is removed from theplacenta, chorion, or amnion-chorion. In some embodiments, some blood isremoved from the placenta, chorion, or amnion-chorion. In someembodiments, the blood is not removed from the placenta, chorion, oramnion-chorion.

In some embodiments, the placenta, chorion, or amnion-chorion iscontacted with a buffer. In some embodiments, the placenta, chorion, oramnion-chorion is contacted with an isotonic buffer. In someembodiments, the placenta, chorion, or amnion-chorion is contacted withsaline, PBS, PBS 1X, Ringer's solution, Hartmann's solution,TRIS-buffered saline, HEPES-buffered saline, EBSS, HBSS, Tyrode's saltSolution, Gey's Balanced Salt Solution, DMEM, EMEM, GMEM, RPMI, or anycombinations thereof.

In some embodiments, the placenta, chorion, or amnion-chorion iscontacted with a suitable antibiotic. In some embodiments, theantibiotic is ciprofloxacin, amphotericin B, penicillin, streptomycin,neomycin or a combination thereof. In some embodiments, the antibioticis ciprofloxacin and amphotericin B. In some embodiments, the antibioticis penicillin, streptomycin, neomycin, and amphotericin B.

In some embodiments, the chorion or amnion-chorion is cut into multiplesheets (e.g., using a scalpel). The size of the sheets depends on thedesired use of the product derived from the chorion or amnion-chorion.In some embodiments, chorion or amnion-chorion is cut into in anysuitable shape (e.g., a square, a circle, a triangle, a rectangle) andinto any suitable sizes.

In some embodiments, the chorion or amnion-chorion sheet is contactedwith a buffer to remove substantially all remaining red blood cells. Insome embodiments, the chorion or amnion-chorion sheet is contacted withan isotonic buffer. In some embodiments, the chorion or amnion-chorionsheet is contacted with saline, PBS, PBS 1X, Ringer's solution,Hartmann's solution, TRIS-buffered saline, HEPES-buffered saline, EBSS,HBSS, Tyrode's salt Solution, Gey's Balanced Salt Solution, DMEM, EMEM,GMEM, RPMI, or any combinations thereof.

In some embodiments, multiple layers of isolated chorion oramnion-chorion are combined to generate a layered chorion oramnion-chorion sheet. In some embodiments, a layered chorion oramnion-chorion sheet comprises at least one layer of chorion and atleast one layer of amnion. The layered chorion or amnion-chorion sheetis any suitable thickness. In some embodiments, the layered chorion oramnion-chorion sheet comprises two, three, four, five, six, seven,eight, nine, or ten layers of isolated chorion or amnion-chorion. Insome embodiments, the layered chorion or amnion-chorion sheet comprisesmore than ten layers of isolated chorion or amnion-chorion.

In some embodiments, the chorion or amnion-chorion sheet is optionallycontacted with a substrate (i.e., a supportive backing). In someembodiments, chorion or amnion-chorion sheet is not contacted with asubstrate. In some embodiments, the chorion or amnion-chorion sheet doesnot require a particular orientation relative to the substrate (i.e.,any side of the chorion or amnion-chorion may be in contact with thesubstrate). In some embodiments, the chorion or amnion-chorion sheet isorientated such that the epithelial layer is in contact with thesubstrate.

In some embodiments, the natural structural integrity of the chorion oramnion-chorion is maintained in the sheet. In some embodiments, thenatural biological activity of the chorion or amnion-chorion ismaintained in the sheet. In some embodiments, the natural structuralintegrity and the natural biological activity of the chorion oramnion-chorion is maintained in the sheet.

In some embodiments, a powder or homogenate is made by: obtainingplacenta comprising chorion, processing the placenta and isolating thechorion (see above for processing and isolation methods), and grinding(or pulverizing) or homogenizing the chorion or amnion-chorion. In someembodiments, the powder or homogenate is made with chorion. In someembodiments, the powder or homogenate is made with amnion-chorion. Insome embodiments, the natural biological activity of the chorion orchorion-amnion (or, any isolated parts thereof) is substantiallymaintained.

In some embodiments, the powder or homogenate is prepared by anysuitable method. In some embodiments, the powder or homogenate isprepared by use of a homogenizer (e.g., an ultrasonic homogenizer),sonicator, pulverizer, Warring blender, grinding mill/jar, bead beater,or any combination thereof.

In some embodiments, the powder or homogenate is prepared by use of agrinding jar. In some embodiments, the chorion or amnion-chorion islyophilized prior to being placed in the grinding jar. A grinding ballis dropped in the grinding jar and the grinding jar is sealed. Thegrinding jar is immersed into liquid nitrogen for 5 min and then placedin a mill and ground at a 30 Hz grinding cycle for 4 min.

In some embodiments, the chorion or amnion-chorion is lyophilized by anysuitable method (e.g., exposure to a liquid gas, placement in afreezer). In some embodiments, the chorion or amnion-chorion (or, anyisolated parts thereof) is placed in the vacuum chamber of alyophilization device until all or substantially all fluid (e.g., water)has been removed. In some embodiments, the placenta (or, any isolatedparts thereof) is lyophilized following freezing (e.g., exposure to atemperature below 0° C., −20° C., −40° C., −50° C., −60° C., −70° C.,−75° C., −80° C., −90° C., −100° C.).

In some embodiments, the natural biological activity of the chorion oramnion-chorion is maintained in the powder or homogenate.

In some embodiments, an extract is made from the chorion oramnion-chorion. In some embodiments, the natural biological activity ofthe chorion or amnion-chorion is maintained in the extract. In someembodiments, a homogenate or powder is made as described above. In someembodiments, the homogenate or powder is centrifuged to generate anextract. Any suitable method of centrifugation may be used. In someembodiments, the extract comprises the supernatant. In some embodiments,the extract comprises the precipitant. In some embodiments, the extractis subject to additional extraction methods (e.g., HABP affinitychromotography, or immuno affinity chromatography).

In some embodiments, the method of making the extract comprises: (a)mixing the homogenate or powder with cold PBS buffer without proteaseinhibitors, to generate a PBS mixture, (b) centrifuging the PBS mixture,and (c) isolating the extract, to generate an isolated extract. In someembodiments, the cold PBS buffer and tissue product are combined in a1:1 ratio. In some embodiments, the PBS mixture is centrifuged at48,000×g 4° C. for 30 min.

In some embodiments, the method of making the extract further comprisespurifying the extract. The number of purification steps depends on thedesired purity. In some embodiments, the purification comprises at least2 rounds of ultracentrifugation. In some embodiments, the purificationcomprises more 2 rounds of ultracentrifugation. In some embodiments, thepurification comprises at least 4 rounds of ultracentrifugation. In someembodiments, the method of purifying the isolated extract comprises: (d)dissolving the isolated extract in CsCl/4M guanidine HCl at the initialdensity of 1.35 g/ml, to generate a CsCl mixture, (e) centrifuging theCsCl mixture at 125,000×g for 48 h at 15° C., to generate a firstpurified extract, (f) extracting the first purified extract anddialyzing it against distilled water to remove CsCl and guanidine HCl,to generate a dialysate. In some embodiments, the method of purifyingthe isolated extract further comprises (g) mixing the dialysate with 3volumes of 95% (v/v) ethanol containing 1.3% (w/v) potassium acetate at0° C. for 1 h, to generate a first dialysate/ethanol mixture, (h)centrifuging the first dialysate/ethanol mixture at 15,000×g, togenerate a second purified extract, and (i) extracting the secondpurified extract. In some embodiments, the method of purifying theisolated extract further comprises: (j) washing the second purifiedextract with ethanol (e.g., 70% ethanol), to generate a second purifiedextract/ethanol mixture; (k) centrifuging the second purifiedextract/ethanol mixture, to generate a third purified extract; and (l)extracting the third purified extract. In some embodiments, the methodof purifying the isolated extract further comprises: (m) washing thethird purified extract with ethanol (e.g., 70% ethanol), to generate athird purified extract/ethanol mixture; (n) centrifuging the thirdpurified extract/ethanol mixture, to generate a forth purified extract;and (o) extracting the forth purified extract. In some embodiments, thepurified extract comprises HC-HA complex.

Storage

In some embodiments, a fetal support tissue product described herein isstored for later use. In some embodiments, storing a fetal supporttissue product described herein does not destroy the natural biologicalactivity of the fetal support tissue product. In some embodiments,storing a fetal support tissue product described herein does not destroythe natural structural integrity of the fetal support tissue product.

Sterilization

In some embodiments, a fetal support tissue product described herein issubject to terminal sterilization by any suitable (e.g., medicallyacceptable) method. In some embodiments, a fetal support tissue productdescribed herein is exposed to gamma radiation for a period of timesufficient to sterilize the fetal support tissue product. In someembodiments, a fetal support tissue product described herein is exposedto gamma radiation at 25 kGy for a period of time sufficient tosterilize the fetal support tissue product. In some embodiments, a fetalsupport tissue product described herein is exposed to gamma radiation at17-30 kGy for a period of time sufficient to sterilize the fetal supporttissue product. In some embodiments, a fetal support tissue productdescribed herein is exposed to an electron beam for a period of timesufficient to sterilize the fetal support tissue product. In someembodiments, a fetal support tissue product described herein is exposedto X-ray radiation for a period of time sufficient to sterilize thefetal support tissue product. In some embodiments, a fetal supporttissue product described herein is exposed to UV radiation for a periodof time sufficient to sterilize the fetal support tissue product.

In some embodiments, the dose range of the gamma-radiation is between 10kGy and 60 kGy, e.g., between 17 kGy and 30 kGy, between 17 kGy and 20kGy, between 20 kGy and 24 kGy, 25 kGy and 30 kGy. In some embodiments,the placental product is radioprotected by exposure to aradioprotectant. In some embodiments, the radioprotectant comprisesglycerol, propylene glycol, trehalose, manitol, DMSO, or a combinationthereof.

In some embodiments, a fetal support tissue product described herein isplaced in a suitable radio-protectant. In some embodiments, theradio-protectant comprises Glycerol, Propylene Glycol, DMSO, Trehalose,Mannitol, or a combination thereof. In some embodiments, theradio-protectant comprises Glycerol, Propylene Glycol, or a combinationthereof.

Storage Medium

In some embodiments, the fetal support tissue product is stored in anysuitable storage medium. In some embodiments, the storage medium isDMEM, Liebowitz's medium, MEM, NCTC, or any combination thereof.

In some embodiments, the storage medium comprises a high oncotic orhyperosmotic agent (also referred to as a “plasma expander”). In someembodiments, the hyperosmotic agent is propylene glycol, glycerol;sugars, such as glucose, sucrose, maltose, dextrose, and the like;dimethyl sulfoxide (DMSO); dimethylamine (DMA); polyvinylpyrrolidone(PVP); sorbitol, glutathione (GSH); ascorbic acid; rosmarinic acid (RO);riboflavin; dextran; albumin; trehalose; mannitol; or any combinationthereof. The hyperosmotic agent maintains an optimal hydration state ofthe fetal support tissue product. Preferably, the hydration of the fetalsupport tissue product is maintained between about 60% and 90% hydrationfor the intended purpose. In some embodiments, the hyperosmotic agentmakes up about 10% to 90% of the storage medium, preferably 10% to about50%, and more preferably about 30% to about 50%. In some embodiments,the fetal support tissue product is stored in 10%, 20%, 30%, 40%, 50%,60%, 70%, 80%, 90% or 100% glycerol. In some embodiments, the fetalsupport tissue product is stored in 10%, 20%, 30%, 40%, 50%, 60%, 70%,80%, 90% or 100% propylene glycol.

In some embodiments, the fetal support tissue product is stored in 50%DMEM+50% Glycerol.

Cryopreservation

In some embodiments, a fetal support tissue product described herein isfrozen for cryopreservation by any suitable method (e.g., exposure to aliquid gas, placement in a freezer, graduated cryopreservation). In someembodiments, cryopreserving a fetal support tissue product describedherein does not destroy the natural biological activity of the fetalsupport tissue product. In some embodiments, cryopreserving a fetalsupport tissue product described herein does not destroy the naturalstructural integrity of the fetal support tissue product.

In some embodiments, a fetal support tissue product described herein isexposed to a liquid gas (e.g., liquid nitrogen or liquid hydrogen). Insome embodiments, the tissue product disclosed herein is exposed toliquid nitrogen. In some embodiments, the fetal support tissue productdisclosed herein does not contact the liquid gas. In some embodiments,the fetal support tissue product disclosed herein is placed in acontainer and the container is contacted with liquid gas. In someembodiments, the fetal support tissue product disclosed herein isexposed to the liquid gas until the fetal support tissue product isfrozen.

In some embodiments, the fetal support tissue product disclosed hereinis frozen by exposure to a temperature below about 0° C. In someembodiments, a fetal support tissue product described herein is frozenby exposure to a liquid gas.

In some embodiments, the fetal support tissue product disclosed hereinis frozen by graduated cryopreservation (e.g., by lowering down thetemperature in a computer-generated program).

In some embodiments, a fetal support tissue product described herein isplaced in a suitable cryo-preservative. In some embodiments, thecryo-preservative comprises Glycerol, Propylene Glycol, DMSO, Trehalose,Mannitol, or a combination thereof. In some embodiments, thecryo-preservative comprises Glycerol, Propylene Glycol, or acombination.

Lyophilization

In some embodiments, the fetal support tissue product disclosed hereinis lyophilized. In some embodiments, lyophilizing a fetal support tissueproduct described herein does not destroy the natural biologicalactivity of the fetal support tissue product. In some embodiments,lyophilizing a fetal support tissue product described herein does notdestroy the natural structural integrity of the fetal support tissueproduct.

In some embodiments, the fetal support tissue product disclosed hereinis lyophilized following freezing. In some embodiments, the fetalsupport tissue product disclosed herein is lyophilized followingfreezing by any suitable method (e.g., exposure to a liquid gas,placement in a freezer).

In some embodiments, a frozen tissue product disclosed herein is placedin the vacuum chamber of a lyophilization device until all orsubstantially all fluid (e.g., water) has been removed.

Rehydration

In some embodiments, a dehydrated or lyophilized product tissue productdisclosed herein is partially or fully rehydrated. In some embodiments,dehydrated or lyophilized product tissue product disclosed hereinrehydrated by contacting the fetal support tissue product with a bufferor with water. In some embodiments, the fetal support tissue product iscontacted with an isotonic buffer. In some embodiments, the fetalsupport tissue product is contacted with saline. In some embodiments,the fetal support tissue product is contacted with PBS. In someembodiments, the fetal support tissue product is contacted with Ringer'ssolution. In some embodiments, the fetal support tissue product iscontacted with Hartmann's solution. In some embodiments, the fetalsupport tissue product is contacted with a TRIS-buffered saline. In someembodiments, the fetal support tissue product is contacted with aHEPES-buffered saline; 50% DMEM+50% Glycerol; 10%, 20%, 30%, 40%, 50%,60%, 70%, 80%, 90% or 100% glycerol; and/or 10%, 20%, 30%, 40%, 50%,60%, 70%, 80%, 90% or 100% propylene glycol.

HC-HA Complex

Disclosed herein, in certain embodiments, are methods of inhibitingosteoclast differentiation in an individual in need thereof, comprisingadministering to the individual a composition comprising HC-HA complex.Disclosed herein, in certain embodiments, are methods of promotingmineralization by osteoblasts in an individual in need thereof,comprising administering to the individual a composition comprisingHC-HA complex. Disclosed herein, in certain embodiments, are methods ofinhibiting bone resorption in an individual in need thereof, comprisingadministering to the individual a composition comprising HC-HA complex.Disclosed herein, in certain embodiments, are methods of inhibiting boneremodeling in an individual in need thereof, comprising administering tothe individual a composition comprising HC-HA complex. Disclosed herein,in certain embodiments, are methods of balancing bone resorption andbone formation, comprising administering to an individual in needthereof a therapeutically-effective amount of a composition comprisingHC-HA complex. Disclosed herein, in certain embodiments, are methods oftreating a disease, disorder, or condition characterized by excessive orundesired osteoclast differentiation, the methods comprisingadministering to an individual in need thereof atherapeutically-effective amount of a composition comprising HC-HAcomplex. Disclosed herein, in certain embodiments, are methods oftreating a disease, disorder, or condition characterized by excessive orundesired bone absorption by osteoclasts, the methods comprisingadministering to an individual in need thereof atherapeutically-effective amount of a composition comprising HC-HAcomplex. Disclosed herein, in certain embodiments, are methods oftreating a disease, disorder, or condition characterized by deficient ordefective bone formation, the methods comprising administering to anindividual in need thereof a therapeutically-effective amount of acomposition comprising HC-HA complex. Disclosed herein, in certainembodiments, are methods of treating arthritis, the methods comprisingadministering to an individual in need thereof atherapeutically-effective amount of a composition comprising HC-HAcomplex. Disclosed herein, in certain embodiments, are methods oftreating osteoporosis, the methods comprising administering to anindividual in need thereof a therapeutically-effective amount of acomposition comprising HC-HA complex. Disclosed herein, in certainembodiments, are methods of treating alveolar bone degradation, themethods comprising administering to an individual in need thereof atherapeutically-effective amount of a composition comprising HC-HAcomplex. Disclosed herein, in certain embodiments, are methods oftreating Paget's disease, the methods comprising administering to anindividual in need thereof a therapeutically-effective amount of acomposition comprising HC-HA complex. Disclosed herein, in certainembodiments, are methods of treating a bone tumor, the methodscomprising administering to an individual in need thereof atherapeutically-effective amount of a composition comprising HC-HAcomplex. In some embodiments of any of the aforementioned methods, theHC-HA complex is isolated from a fetal support tissue (e.g., UCAM, PAM,umbilical cord, placenta, chorion, amnion-chorion, or any combinationsthereof) and purified. In some embodiments of any of the aforementionedmethods, the HC-HA complex is obtained by a process comprising (a)providing a reaction mixture comprising: (i) HA (e.g., HMW HA); (ii)IαI, wherein the IαI is optionally in serum or isolated from serum;(iii); (iii) TSG-6, wherein the TSG-6 is optionally recombinant; and(iv) PTX3, wherein the PTX3 is optionally recombinant; wherein at leastone of HA, IαI, TSG-6, or PTX3 is optionally generated by a plurality ofcells present in the reaction mixture; (b) incubating the reactionmixture for a period of time sufficient to produce HC-HA complex; and(c) isolating and purifying the HC-HA complex.

Manufacturing HC-HA

In some embodiments of any of the aforementioned methods, the HC-HAcomplex is obtained by a process comprising (a) providing a reactionmixture comprising: (i) HA (e.g., HMW HA); (ii) IαI, wherein the IαI isoptionally in serum or isolated from serum; (iii); (iii) TSG-6, whereinthe TSG-6 is optionally recombinant; and (iv) PTX3, wherein the PTX3 isoptionally recombinant; wherein at least one of HA, IαI, TSG-6, or PTX3is optionally generated by a plurality of cells present in the reactionmixture; (b) incubating the reaction mixture for a period of timesufficient to produce HC-HA complex; and (c) isolating and purifying theHC-HA complex. In some embodiments, the contacting occurs for at least 6hours, at least 12 hours, at least 24 hours, at least 36 hours, at least48 hours, at least 60 hours, or at least 72 hours.

In some embodiments, the method further comprises immobilizing HA (e.g.,HMW HA) to a stationary support (e.g., by cross-linking) In someembodiments, the stationary support comprising HA (e.g., HMW HA) iscontacted with IαI (e.g., IαI purified from serum, IαI in serum), TSG-6(or, recombinant TSG-6), and PTX3 (or, recombinant PTX3). In someembodiments, the contacting occurs for at least 6 hours, at least 12hours, at least 24 hours, at least 36 hours, at least 48 hours, at least60 hours, or at least 72 hours. In some embodiments, the stationarysupport is washed to remove any unbound components.

In some embodiments, the method further comprises HA binding protein(HABP). In some embodiments, HABP is affixed to a stationary support(e.g., by cross-linking). In some embodiments, the stationary supportcomprising HABP is contacted with HA (e.g., HMW HA), IαI, TSG-6 (or,recombinant TSG-6), and PTX3 (or, recombinant PTX3). In someembodiments, the contacting occurs for at least 6 hours, at least 12hours, at least 24 hours, at least 36 hours, at least 48 hours, at least60 hours, or at least 72 hours. In some embodiments, the stationarysupport is washed to remove any unbound components.

In some embodiments, IαI is isolated from serum. In some embodiments,IαI is in serum. In some embodiments, IαI is isolated from a cell orplurality of cells.

In some embodiments, TSG6 is isolated from a cell or a plurality ofcells (e.g., a tissue extract). In some embodiments, TSG6 is notisolated from a cell or a plurality of cells (e.g., a tissue extract).In some embodiments, TSG6 is prepared by recombinant technology.

In some embodiments, PTX3 is isolated from a cell or a plurality ofcells (e.g., a tissue extract). In some embodiments, PTX3 is notisolated from a cell or a plurality of cells (e.g., a tissue extract).In some embodiments, PTX3 is prepared by recombinant technology.

Purification of HC-HA

In some embodiments, the HC-HA is purified by any suitable method. Insome embodiments, the HC-HA complex is purified by centrifugation (e.g.,ultracentrifugation, gradient centrifugation), chromatography (e.g., ionexchange, affinity, size exclusion, and hydroxyapatite chromatography),gel filtration, or differential solubility, ethanol precipitation or byany other available technique for the purification of proteins (See,e.g., Scopes, Protein Purification Principles and Practice 2nd Edition,Springer-Verlag, New York, 1987; Higgins, S. J. and Hames, B. D. (eds.),Protein Expression: A Practical Approach, Oxford Univ Press, 1999; andDeutscher, M. P., Simon, M. I., Abelson, J. N. (eds.), Guide to ProteinPurification: Methods in Enzymology (Methods in Enzymology Series, Vol182), Academic Press, 1997, all incorporated herein by reference).

In some embodiments, the HC-HA complex is purified byultracentrifugation. The number of purification steps depends on thedesired purity. In some embodiments, the purification comprises at least2 rounds of ultracentrifugation. In some embodiments, the purificationcomprises more 2 rounds of ultracentrifugation. In some embodiments, thepurification comprises at least 4 rounds of ultracentrifugation. In someembodiments, the method of purifying the isolated extract comprises: (d)dissolving the isolated extract in CsCl/4M guanidine HCl at the initialdensity of 1.35 g/ml, to generate a CsCl mixture, (e) centrifuging theCsCl mixture at 125,000×g for 48 h at 15° C., to generate a firstpurified extract, (f) extracting the first purified extract anddialyzing it against distilled water to remove CsCl and guanidine HCl,to generate a dialysate. In some embodiments, the method of purifyingthe isolated extract further comprises (g) mixing the dialysate with 3volumes of 95% (v/v) ethanol containing 1.3% (w/v) potassium acetate at0° C. for 1 h, to generate a first dialysate/ethanol mixture, (h)centrifuging the first dialysate/ethanol mixture at 15,000×g, togenerate a second purified extract, and (i) extracting the secondpurified extract. In some embodiments, the method of purifying theisolated extract further comprises: (j) washing the second purifiedextract with ethanol (e.g., 70% ethanol), to generate a second purifiedextract/ethanol mixture; (k) centrifuging the second purifiedextract/ethanol mixture, to generate a third purified extract; and (l)extracting the third purified extract. In some embodiments, the methodof purifying the isolated extract further comprises: (m) washing thethird purified extract with ethanol (e.g., 70% ethanol), to generate athird purified extract/ethanol mixture; (n) centrifuging the thirdpurified extract/ethanol mixture, to generate a forth purified extract;and (o) extracting the forth purified extract. The preferredpurification method comprises four rounds of ultracentrifugation.

In some embodiments, the HC-HA complex is purified by immunoaffinitychromatography. In some embodiments, anti HC1 antibodies, anti-HC2antibodies, or both are generated and affixed to a stationary support.In some embodiments, the unpurified HC-HA complex (i.e., the mobilephase) is passed over the support. In certain instances, the HC-HAcomplex binds to the antibodies (e.g., via interaction of (a) an HC1antibody and HC1, (b) an HC2 antibody and HC2, or (c) both). In someembodiments the support is washed (e.g., with PBS) to remove any unboundor loosely bound molecules. In some embodiments, the support is thenwashed with a solution that enables elution of the HC-HA complex fromthe support (e.g., 1% SDS, 6M guanidine-HC1, or 8M urea).

In some embodiments, the HC-HA complex is purified by affinitychromatography. In some embodiments, HABP is generated and affixed to astationary support. In some embodiments, the unpurified HC-HA complex(i.e., the mobile phase) is passed over the support. In certaininstances, the HC-HA complex binds to the HABP. In some embodiments thesupport is washed (e.g., with PBS) to remove any unbound or looselybound molecules. In some embodiments, the support is then washed with asolution that enables elution of the HC-HA complex from the support.

In some embodiments, the HC-HA complex is purified by a combination ofHABP affinity chromotography, and immunoaffinity chromatography usinganti HC1 antibodies, anti-HC 2 antibodies, or both.

Bioreactor

In some embodiments, the HC-HA complex is made by use of live cells. Insome embodiments, the method comprises contacting (a) hyaluronan (HA);(b) IαI; (c) TSG-6; and (d) PTX3; wherein one or more components isgenerated or expressed by a plurality of cells in a bioreactor.

In some embodiments, the method comprises HA that is obtained from acommercial supplier. In some embodiments, the method comprises HA thatis generated by a plurality of cells in a bioreactor. In someembodiments, the plurality of cells constitutively generate HA. In someembodiments, the plurality of cells constitutively expresses HAS1, HAS2,HAS3, or a combination thereof. In some embodiments, the plurality ofcells are contacted with at least one factor known to upregulate HAS1,HAS2, HAS3, or a combination thereof.

In some embodiments, the method comprises IαI isolated from serum. Insome embodiments, the method comprises IαI that is not isolated fromserum. In some embodiments, the method comprises IαI that is expressedby a cell or a plurality of cells in a bioreactor. In some embodiments,the plurality of cells constitutively expresses IαI.

In some embodiments, the method comprises TSG-6 that is isolated from acell or a plurality of cells (e.g., a tissue extract). In someembodiments, the method comprises TSG-6 that is not isolated from a cellor a plurality of cells (e.g., a tissue extract). In some embodiments,the method comprises TSG-6 that is expressed by a cell or a plurality ofcells in a bioreactor. In some embodiments, the cell or plurality ofcells constitutively generates TSG-6. In some embodiments, the cell orplurality of cells constitutively expresses TSG-6. In some embodiments,the cell or plurality of cells is contacted with at least one factorknown to upregulate TSG-6.

In some embodiments, the method comprises PTX3 that is isolated from acell or a plurality of cells (e.g., a tissue extract). In someembodiments, the method comprises PTX3 that is not isolated from a cellor a plurality of cells (e.g., a tissue extract). In some embodiments,the method comprises PTX3 that is expressed by a cell or a plurality ofcells in a bioreactor. In some embodiments, the cell or plurality ofcells constitutively generates PTX3. In some embodiments, the cell orplurality of cells constitutively expresses PTX3. In some embodiments,the cell or plurality of cells is contacted with at least one factorknown to upregulate expression of PTX3.

In some embodiments, a cell that constitutively (a) expresses IαI; (b)expresses TSG-6; or (c) PTX3, is generated by any suitable method. Insome embodiments, a cell that constitutively (a) expresses IαI; (b)expresses TSG-6; or (c) expresses PTX3 is generated by introducing pointmutations into a gene encoding (a); (b) TSG-6; or (c) PTX3. In someembodiments, the mutations are substitution mutations, deletionmutations, or insertion mutations.

In some embodiments, a cell that constitutively generates HA is producedby any suitable method. In some embodiments, a cell that constitutivelygenerates HA is produced by introducing point mutations into a geneencoding HAS1, HAS2, HAS3, or a combination thereof. In someembodiments, the mutations are substitution mutations, deletionmutations, or insertion mutations. In some embodiments, a cell thatconstitutively generates HA is produced by contacting the cell with atleast one factor known to upregulate HAS1, HAS2, HAS3, or a combinationthereof.

Pharmaceutical Compositions

Disclosed herein, in certain embodiments, are methods of inhibitingosteoclast differentiation in an individual in need thereof, comprisingadministering to the individual a composition comprising product fetalsupport tissue product or an extract thereof. Disclosed herein, incertain embodiments, are methods of promoting mineralization byosteoblasts in an individual in need thereof, comprising administeringto the individual a composition comprising a fetal support tissueproduct or an extract thereof. In some embodiments, the fetal supporttissue product is substantially isolated placental amniotic membrane(PAM), umbilical cord amniotic membrane (UCAM), placenta, umbilicalcord, chorion, amnion-chorion, or any combinations thereof. Disclosedherein, in certain embodiments, are methods of inhibiting boneresorption in an individual in need thereof, comprising administering tothe individual a composition comprising a fetal support tissue productor an extract thereof. Disclosed herein, in certain embodiments, aremethods of inhibiting bone remodeling in an individual in need thereof,comprising administering to the individual a composition comprising afetal support tissue product or an extract thereof. Disclosed herein, incertain embodiments, are methods of balancing bone resorption and boneformation, comprising administering to an individual in need thereof atherapeutically-effective amount of a composition comprising a fetalsupport tissue product or an extract thereof. Disclosed herein, incertain embodiments, are methods of treating a disease, disorder, orcondition characterized by excessive or undesired osteoclastdifferentiation, the methods comprising administering to an individualin need thereof a therapeutically-effective amount of a compositioncomprising a fetal support tissue product or an extract thereof.Disclosed herein, in certain embodiments, are methods of treating adisease, disorder, or condition characterized by excessive or undesiredbone absorption by osteoclasts, the methods comprising administering toan individual in need thereof a therapeutically-effective amount of acomposition comprising a fetal support tissue product or an extractthereof. Disclosed herein, in certain embodiments, are methods oftreating a disease, disorder, or condition characterized by deficient ordefective bone formation, the methods comprising administering to anindividual in need thereof a therapeutically-effective amount of acomposition comprising a fetal support tissue product or an extractthereof. Disclosed herein, in certain embodiments, are methods oftreating arthritis, the methods comprising administering to anindividual in need thereof a therapeutically-effective amount of acomposition comprising a fetal support tissue product or an extractthereof. Disclosed herein, in certain embodiments, are methods oftreating osteoporosis, the methods comprising administering to anindividual in need thereof a therapeutically-effective amount of acomposition comprising a fetal support tissue product or an extractthereof. Disclosed herein, in certain embodiments, are methods oftreating alveolar bone degradation, the methods comprising administeringto an individual in need thereof a therapeutically-effective amount of acomposition comprising a fetal support tissue product or an extractthereof. Disclosed herein, in certain embodiments, are methods oftreating Paget's disease, the methods comprising administering to anindividual in need thereof a therapeutically-effective amount of acomposition comprising a fetal support tissue product or an extractthereof. Disclosed herein, in certain embodiments, are methods oftreating a bone tumor, the methods comprising administering to anindividual in need thereof a therapeutically-effective amount of acomposition comprising a fetal support tissue product or an extractthereof. In some embodiments, the compositions further comprisepharmaceutically acceptable diluent(s), excipient(s), or carrier(s). Insome embodiments, the compositions further comprise an adhesive. In yetother embodiments, the compositions further comprise a penetrationenhancer. In some embodiments, the compositions comprise a viscosityenhancing agent.

In other embodiments, the pharmaceutical compositions further comprise afurther therapeutic agent. For a non-limiting disclosure of furthertherapeutic agents see the Combination Therapies section herein.

The pharmaceutical compositions described herein can be administered toa subject by multiple administration routes, including but not limitedto, oral, parenteral (e.g., intravenous, subcutaneous, intramuscular),intranasal, buccal, topical, rectal, or transdermal administrationroutes. The pharmaceutical formulations described herein include, butare not limited to, aqueous liquid dispersions, self-emulsifyingdispersions, solid solutions, liposomal dispersions, aerosols, soliddosage forms, powders, immediate release formulations, controlledrelease formulations, fast melt formulations, tablets, capsules, pills,delayed release formulations, extended release formulations, pulsatilerelease formulations, multiparticulate formulations, and mixed immediateand controlled release formulations.

In some embodiments, the compositions described herein are formulated assolutions. In some embodiments, the compositions are formulated as gels.In some embodiments, the compositions described herein are formulated asliposomes. In some embodiments, the compositions described herein areformulated as foams. In some embodiments, the compositions describedherein are formulated as paints. In some embodiments, the compositionsdescribed herein are formulated as in situ forming spongy materials.

In some embodiments, the compositions disclosed herein are formulatedfor immediate release. In some embodiments, the compositions areformulated for controlled release (e.g., delayed release or sustainedrelease). In some embodiments, the compositions are formulated forimmediate release and controlled release. As used herein, “controlledrelease” includes delayed release, sustained release, extended release,variable release, pulsatile release and bi-modal release. Thecontrolled-release aspect of the compositions described herein isimparted by any suitable excipients. By way of example only, suchexcipients include polymers and, viscosity enhancing agents.

In some embodiments, the compositions are biodegradable.

Pharmaceutical compositions may be formulated in a conventional mannerusing one or more physiologically acceptable carriers includingexcipients and auxiliaries which facilitate processing of the activecompounds into preparations which can be used pharmaceutically. Properformulation is dependent upon the route of administration chosen. Any ofthe well-known techniques, carriers, and excipients may be used assuitable and as understood in the art. A summary of pharmaceuticalcompositions described herein may be found, for example, in Remington:The Science and Practice of Pharmacy, Nineteenth Ed (Easton, Pa.: MackPublishing Company, 1995); Hoover, John E., Remington's PharmaceuticalSciences, Mack Publishing Co., Easton, Pa. 1975; Liberman, H. A. andLachman, L., Eds., Pharmaceutical Dosage Forms, Marcel Decker, New York,N.Y., 1980; and Pharmaceutical Dosage Forms and Drug Delivery Systems,Seventh Ed. (Lippincott Williams & Wilkins 1999), herein incorporated byreference in their entirety.

Pharmaceutical compositions including a compound described herein may bemanufactured in a conventional manner, such as, by way of example only,by means of conventional mixing, dissolving, granulating, dragee-making,levigating, emulsifying, encapsulating, entrapping or compressionprocesses.

Excipients

In some embodiments, the compositions further comprise pharmaceuticallyacceptable diluent(s), excipient(s), or carrier(s).

In some embodiments, the compositions further comprise an antifoamingagent. “Antifoaming agents” reduce foaming during processing which canresult in coagulation of aqueous dispersions, bubbles in the finishedfilm, or generally impair processing. Exemplary anti-foaming agentsinclude silicon emulsions or sorbitan sesquoleate.

In some embodiments, the compositions further comprise an antioxidant.“Antioxidants” include, for example, butylated hydroxytoluene (BHT),sodium ascorbate, ascorbic acid, sodium metabisulfite and tocopherol. Incertain embodiments, antioxidants enhance chemical stability whererequired.

In some embodiments, the compositions further comprise a binder.“Binders” impart cohesive qualities and include, e.g., alginic acid andsalts thereof; cellulose derivatives such as carboxymethylcellulose,methylcellulose (e.g., Methocel®), hydroxypropylmethylcellulose,hydroxyethylcellulose, hydroxypropylcellulose (e.g., Klucel®),ethylcellulose (e.g., Ethocel®), and microcrystalline cellulose (e.g.,Avicel®); microcrystalline dextrose; amylose; magnesium aluminumsilicate; polysaccharide acids; bentonites; gelatin;polyvinylpyrrolidone/vinyl acetate copolymer; crosspovidone; povidone;starch; pregelatinized starch; tragacanth, dextrin, a sugar, such assucrose (e.g., Dipac®), glucose, dextrose, molasses, mannitol, sorbitol,xylitol (e.g., Xylitab®), and lactose; a natural or synthetic gum suchas acacia, tragacanth, ghatti gum, mucilage of isapol husks,polyvinylpyrrolidone (e.g., Polyvidone® CL, Kollidon® CL, Polyplasdone®XL-10), larch arabogalactan, Veegum®, polyethylene glycol, waxes, sodiumalginate, and the like.

In some embodiments, the compositions described herein further comprisebuffering agents, for example to impart a physiological acceptable pH.Suitable buffering agent include acids such as acetic, boric, citric,lactic, phosphoric and hydrochloric acids; bases such as sodiumhydroxide, sodium phosphate, sodium borate, sodium citrate, sodiumacetate, sodium lactate and tris-hydroxymethylaminomethane; and bufferssuch as citrate/dextrose, sodium bicarbonate and ammonium chloride. Suchacids, bases and buffers are included in an amount required to maintainpH of the composition in an acceptable range.

In some embodiments, the compositions further comprise a carrier. A“carrier” or “carrier materials” include any commonly used excipients inpharmaceutics and should be selected on the basis of compatibility withcompounds disclosed herein, such as, compounds of any of Formula D andthe second agent, and the release profile properties of the desireddosage form. Exemplary carrier materials include, e.g., binders,suspending agents, disintegration agents, filling agents, surfactants,solubilizers, stabilizers, lubricants, wetting agents, diluents, and thelike. “Pharmaceutically compatible carrier materials” may include, butare not limited to, acacia, gelatin, colloidal silicon dioxide, calciumglycerophosphate, calcium lactate, maltodextrin, glycerine, magnesiumsilicate, polyvinylpyrrollidone (PVP), cholesterol, cholesterol esters,sodium caseinate, soy lecithin, taurocholic acid, phosphotidylcholine,sodium chloride, tricalcium phosphate, dipotassium phosphate, celluloseand cellulose conjugates, sugars sodium stearoyl lactylate, carrageenan,monoglyceride, diglyceride, pregelatinized starch, and the like. See,e.g., Remington: The Science and Practice of Pharmacy, Nineteenth Ed(Easton, Pa.: Mack Publishing Company, 1995); Hoover, John E.,Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa.1975; Liberman, H. A. and Lachman, L., Eds., Pharmaceutical DosageForms, Marcel Decker, New York, N.Y., 1980; and Pharmaceutical DosageForms and Drug Delivery Systems, Seventh Ed. (Lippincott Williams &Wilkins 1999).

In some embodiments, the compositions further comprise a dispersingagent and/or viscosity modulating agents. “Dispersing agents,” and/or“viscosity modulating agents” include materials that control thediffusion and homogeneity of a drug through liquid media or agranulation method or blend method. In some embodiments, these agentsalso facilitate the effectiveness of a coating or eroding matrix.Exemplary diffusion facilitators/dispersing agents include, e.g.,hydrophilic polymers, electrolytes, Tween® 60 or 80, PEG,polyvinylpyrrolidone (PVP; commercially known as Plasdone®), and thecarbohydrate-based dispersing agents such as, for example, hydroxypropylcelluloses (e.g., HPC, HPC-SL, and HPC-L), hydroxypropylmethylcelluloses (e.g., HPMC K100, HPMC K4M, HPMC K15M, and HPMC K100M),carboxymethylcellulose sodium, methylcellulose, hydroxyethylcellulose,hydroxypropylcellulose, hydroxypropylmethylcellulose phthalate,hydroxypropylmethylcellulose acetate stearate (HPMCAS), noncrystallinecellulose, magnesium aluminum silicate, triethanolamine, polyvinylalcohol (PVA), vinyl pyrrolidone/vinyl acetate copolymer (S630),4-(1,1,3,3-tetramethylbutyl)-phenolpolymer with ethylene oxide andformaldehyde (also known as tyloxapol), poloxamers (e.g., PluronicsF68®, F88®, and F108®, which are block copolymers of ethylene oxide andpropylene oxide); and poloxamines (e.g., Tetronic 908®, also known asPoloxamine 908®, which is a tetrafunctional block copolymer derived fromsequential addition of propylene oxide and ethylene oxide toethylenediamine (BASF Corporation, Parsippany, N.J.)),polyvinylpyrrolidone K12, polyvinylpyrrolidone K17, polyvinylpyrrolidoneK25, or polyvinylpyrrolidone K30, polyvinylpyrrolidone/vinyl acetatecopolymer (S-630), polyethylene glycol, e.g., the polyethylene glycolcan have a molecular weight of about 300 to about 6000, or about 3350 toabout 4000, or about 7000 to about 5400, sodium carboxymethylcellulose,methylcellulose, polysorbate-80, sodium alginate, gums, such as, e.g.,gum tragacanth and gum acacia, guar gum, xanthans, including xanthangum, sugars, cellulosics, such as, e.g., sodium carboxymethylcellulose,methylcellulose, sodium carboxymethylcellulose, polysorbate-80, sodiumalginate, polyethoxylated sorbitan monolaurate, polyethoxylated sorbitanmonolaurate, povidone, carbomers, polyvinyl alcohol (PVA), alginates,chitosans and combinations thereof. Plasticizers such as cellulose ortriethyl cellulose can also be used as dispersing agents. Dispersingagents particularly useful in liposomal dispersions and self-emulsifyingdispersions are dimyristoyl phosphatidyl choline, natural phosphatidylcholine from eggs, natural phosphatidyl glycerol from eggs, cholesteroland isopropyl myristate.

In some embodiments, the compositions further comprise a diluent. Theterm “diluent” refers to chemical compounds that are used to dilute thecompound of interest prior to delivery. Diluents can also be used tostabilize compounds because they can provide a more stable environment.Salts dissolved in buffered solutions (which also can provide pH controlor maintenance) are utilized as diluents in the art, including, but notlimited to a phosphate buffered saline solution. In certain embodiments,diluents increase bulk of the composition to facilitate compression orcreate sufficient bulk for homogenous blend for capsule filling. Suchcompounds include e.g., lactose, starch, mannitol, sorbitol, dextrose,microcrystalline cellulose such as Avicel®; dibasic calcium phosphate,dicalcium phosphate dihydrate; tricalcium phosphate, calcium phosphate;anhydrous lactose, spray-dried lactose; pregelatinized starch,compressible sugar, such as Di-Pac® (Amstar); mannitol,hydroxypropylmethylcellulose, hydroxypropylmethylcellulose acetatestearate, sucrose-based diluents, confectioner's sugar; monobasiccalcium sulfate monohydrate, calcium sulfate dihydrate; calcium lactatetrihydrate, dextrates; hydrolyzed cereal solids, amylose; powderedcellulose, calcium carbonate; glycine, kaolin; mannitol, sodiumchloride; inositol, bentonite, and the like.

In some embodiments, the compositions further comprise a disintegrant.The term “disintegrate” includes both the dissolution and dispersion ofthe dosage form when contacted with gastrointestinal fluid.“Disintegration agents or disintegrants” facilitate the breakup ordisintegration of a substance. Examples of disintegration agents includea starch, e.g., a natural starch such as corn starch or potato starch, apregelatinized starch such as National 1551 or Amijel®, or sodium starchglycolate such as Promogel® or Explotab®, a cellulose such as a woodproduct, methylcrystalline cellulose, e.g., Avicel®, Avicel® PH101,Avicel® PH102, Avicel® PH105, Elcema® P100, Emcocel®, Vivacel®, MingTia®, and Solka-Floc®, methylcellulose, croscarmellose, or across-linked cellulose, such as cross-linked sodiumcarboxymethylcellulose (Ac-Di-Sol®), cross-linkedcarboxymethylcellulose, or cross-linked croscarmellose, a cross-linkedstarch such as sodium starch glycolate, a cross-linked polymer such ascrosspovidone, a cross-linked polyvinylpyrrolidone, alginate such asalginic acid or a salt of alginic acid such as sodium alginate, a claysuch as Veegum® HV (magnesium aluminum silicate), a gum such as agar,guar, locust bean, Karaya, pectin, or tragacanth, sodium starchglycolate, bentonite, a natural sponge, a surfactant, a resin such as acation-exchange resin, citrus pulp, sodium lauryl sulfate, sodium laurylsulfate in combination starch, and the like.

In some embodiments, the compositions further comprise a preservative.Suitable preservatives include mercury-containing substances such asmerfen and thiomersal; stabilized chlorine dioxide; and quaternaryammonium compounds such as benzalkonium chloride, cetyltrimethylammoniumbromide and cetylpyridinium chloride.

In some embodiments, the compositions described herein further comprisesalts. In other embodiments, compositions described herein furthercomprise one or more salts in an amount sufficient to impart aphysiologically acceptable osmolarity. Such salts include those havingsodium, potassium or ammonium cations and chloride, citrate, ascorbate,borate, phosphate, bicarbonate, sulfate, thiosulfate or bisulfiteanions; suitable salts include sodium chloride, potassium chloride,sodium thiosulfate, sodium bisulfite and ammonium sulfate.

In some embodiments, the compositions further comprise a solubilizer.“Solubilizers” include compounds such as triacetin, triethylcitrate,ethyl oleate, ethyl caprylate, sodium lauryl sulfate, sodium doccusate,vitamin E TPGS, dimethylacetamide, N-methylpyrrolidone,N-hydroxyethylpyrrolidone, polyvinylpyrrolidone, hydroxypropylmethylcellulose, hydroxypropyl cyclodextrins, ethanol, n-butanol, isopropylalcohol, cholesterol, bile salts, polyethylene glycol 200-600,glycofurol, transcutol, propylene glycol, and dimethyl isosorbide andthe like.

“Stabilizers” include compounds such as any antioxidation agents,buffers, acids, preservatives and the like.

In some embodiments, the compositions further comprise a suspendingagent. “Suspending agents” include compounds such aspolyvinylpyrrolidone, e.g., polyvinylpyrrolidone K12,polyvinylpyrrolidone K17, polyvinylpyrrolidone K25, orpolyvinylpyrrolidone K30, vinyl pyrrolidone/vinyl acetate copolymer(S630), polyethylene glycol, e.g., the polyethylene glycol can have amolecular weight of about 300 to about 6000, or about 3350 to about4000, or about 7000 to about 5400, sodium carboxymethylcellulose,methylcellulose, hydroxypropylmethylcellulose, hydroxymethylcelluloseacetate stearate, polysorbate-80, hydroxyethylcellulose, sodiumalginate, gums, such as, e.g., gum tragacanth and gum acacia, guar gum,xanthans, including xanthan gum, sugars, cellulosics, such as, e.g.,sodium carboxymethylcellulose, methylcellulose, sodiumcarboxymethylcellulose, hydroxypropylmethylcellulose,hydroxyethylcellulose, polysorbate-80, sodium alginate, polyethoxylatedsorbitan monolaurate, polyethoxylated sorbitan monolaurate, povidone andthe like.

In some embodiments, the compositions further comprise a surfactant.“Surfactants” include compounds such as sodium lauryl sulfate, sodiumdocusate, Tween 60 or 80, triacetin, vitamin E TPGS, sorbitanmonooleate, polyoxyethylene sorbitan monooleate, polysorbates,polaxomers, bile salts, glyceryl monostearate, copolymers of ethyleneoxide and propylene oxide, e.g., Pluronic® (BASF), and the like. Someother surfactants include polyoxyethylene fatty acid glycerides andvegetable oils, e.g., polyoxyethylene (60) hydrogenated castor oil; andpolyoxyethylene alkylethers and alkylphenyl ethers, e.g., octoxynol 10,octoxynol 40. In some embodiments, surfactants may be included toenhance physical stability or for other purposes.

In some embodiments, the compositions further comprise a viscosityenhancing agent. “Viscosity enhancing agents” include, e.g., methylcellulose, xanthan gum, carboxymethyl cellulose, hydroxypropylcellulose, hydroxypropylmethyl cellulose, hydroxypropylmethyl celluloseacetate stearate, hydroxypropylmethyl cellulose phthalate, carbomer,polyvinyl alcohol, alginates, acacia, chitosans and combinationsthereof.

In some embodiments, the compositions further comprise a wetting agent.“Wetting agents” include compounds such as oleic acid, glycerylmonostearate, sorbitan monooleate, sorbitan monolaurate, triethanolamineoleate, polyoxyethylene sorbitan monooleate, polyoxyethylene sorbitanmonolaurate, sodium docusate, sodium oleate, sodium lauryl sulfate,sodium doccusate, triacetin, Tween 80, vitamin E TPGS, ammonium saltsand the like.

Dosage Forms

The compositions described herein e.g., HC-HA, fetal support tissueextracts, fetal support powders and homogenates) can be formulated foradministration to a subject via any conventional means including, butnot limited to, oral, parenteral (e.g., intravenous, subcutaneous, orintramuscular), buccal, intranasal, rectal or transdermal administrationroutes. As used herein, the term “subject” is used to mean an animal,preferably a mammal, including a human or non-human. The terms patientand subject may be used interchangeably.

Moreover, the pharmaceutical compositions (e.g., HC-HA, fetal supporttissue extracts, fetal support powders and homogenates) disclosed hereincan be formulated into any suitable dosage form, including but notlimited to, aqueous oral dispersions, liquids, gels, syrups, elixirs,slurries, suspensions and the like, for oral ingestion by a patient tobe treated, solid oral dosage forms, aerosols, controlled releaseformulations, fast melt formulations, effervescent formulations,lyophilized formulations, tablets, powders, pills, dragees, capsules,delayed release formulations, extended release formulations, pulsatilerelease formulations, multiparticulate formulations, and mixed immediaterelease and controlled release formulations.

Solutions

In some embodiments, the compositions (e.g., HC-HA, fetal support tissueextracts, fetal support powders and homogenates) disclosed herein reformulated as solutions. Solutions include aqueous solutions,dispersions, and emulsions. See, e.g., Singh et al., Encyclopedia ofPharmaceutical Technology, 2^(nd) Ed., pp. 754-757 (2002).

The aqueous suspensions and dispersions described herein can remain in ahomogenous state, as defined in The USP Pharmacists' Pharmacopeia (2005edition, chapter 905), for at least 4 hours. The homogeneity should bedetermined by a sampling method consistent with regard to determininghomogeneity of the entire composition. In one embodiment, an aqueoussuspension can be re-suspended into a homogenous suspension by physicalagitation lasting less than 1 minute. In another embodiment, an aqueoussuspension can be re-suspended into a homogenous suspension by physicalagitation lasting less than 45 seconds. In yet another embodiment, anaqueous suspension can be re-suspended into a homogenous suspension byphysical agitation lasting less than 30 seconds. In still anotherembodiment, no agitation is necessary to maintain a homogeneous aqueousdispersion.

Examples of disintegrating agents for use in the aqueous suspensions anddispersions include, but are not limited to, a starch, e.g., a naturalstarch such as corn starch or potato starch, a pregelatinized starchsuch as National 1551 or Amijel®, or sodium starch glycolate such asPromogel® or Explotab®; a cellulose such as a wood product,methylcrystalline cellulose, e.g., Avicel®, Avicel® PH101, Avicer PH102,Avicel® PH105, Elcema® P100, Emcocel®, Vivacel®, Ming Tia®, andSolka-Floc®, methylcellulose, croscarmellose, or a cross-linkedcellulose, such as cross-linked sodium carboxymethylcellulose(Ac-Di-Sol®), cross-linked carboxymethylcellulose, or cross-linkedcroscarmellose; a cross-linked starch such as sodium starch glycolate; across-linked polymer such as crospovidone; a cross-linkedpolyvinylpyrrolidone; alginate such as alginic acid or a salt of alginicacid such as sodium alginate; a clay such as Veegum® HV (magnesiumaluminum silicate); a gum such as agar, guar, locust bean, Karaya,pectin, or tragacanth; sodium starch glycolate; bentonite; a naturalsponge; a surfactant; a resin such as a cation-exchange resin; citruspulp; sodium lauryl sulfate; sodium lauryl sulfate in combinationstarch; and the like.

In some embodiments, the dispersing agents suitable for the aqueoussuspensions and dispersions described herein are known in the art andinclude, for example, hydrophilic polymers, electrolytes, Tween® 60 or80, PEG, polyvinylpyrrolidone (PVP; commercially known as Plasdone®),and the carbohydrate-based dispersing agents such as, for example,hydroxypropylcellulose and hydroxypropyl cellulose ethers (e.g., HPC,HPC-SL, and HPC-L), hydroxypropyl methylcellulose and hydroxypropylmethylcellulose ethers (e.g. HPMC K100, HPMC K4M, HPMC K15M, and HPMCK100M), carboxymethylcellulose sodium, methylcellulose,hydroxyethylcellulose, hydroxypropylmethyl-cellulose phthalate,hydroxypropylmethyl-cellulose acetate stearate, noncrystallinecellulose, magnesium aluminum silicate, triethanolamine, polyvinylalcohol (PVA), polyvinylpyrrolidone/vinyl acetate copolymer (Plasdone®,e.g., S-630), 4-(1,1,3,3-tetramethylbutyl)-phenolpolymer with ethyleneoxide and formaldehyde (also known as tyloxapol), poloxamers (e.g.,Pluronics F68®, F88®, and F108®, which are block copolymers of ethyleneoxide and propylene oxide); and poloxamines (e.g., Tetronic 908®, alsoknown as Poloxamine 908®, which is a tetrafunctional block copolymerderived from sequential addition of propylene oxide and ethylene oxideto ethylenediamine (BASF Corporation, Parsippany, N.J.)). In otherembodiments, the dispersing agent is selected from a group notcomprising one of the following agents: hydrophilic polymers;electrolytes; Tween® 60 or 80; PEG; polyvinylpyrrolidone (PVP);hydroxypropylcellulose and hydroxypropyl cellulose ethers (e.g., HPC,HPC-SL, and HPC-L); hydroxypropyl methylcellulose and hydroxypropylmethylcellulose ethers (e.g. HPMC K100, HPMC K4M, HPMC K15M, HPMC K100M,and Pharmacoat® USP 2910 (Shin-Etsu)); carboxymethylcellulose sodium;methylcellulose; hydroxyethylcellulose; hydroxypropylmethylcellulosephthalate; hydroxypropylmethyl-cellulose acetate stearate;non-crystalline cellulose; magnesium aluminum silicate; triethanolamine;polyvinyl alcohol (PVA); 4-(1,1,3,3-tetramethylbutyl)-phenolpolymer withethylene oxide and formaldehyde; poloxamers (e.g., Pluronics F68®, F88®,and F108®, which are block copolymers of ethylene oxide and propyleneoxide); or poloxamines (e.g., Tetronic 908®, also known as Poloxamine908®).

Wetting agents suitable for the aqueous suspensions and dispersionsdescribed herein are known in the art and include, but are not limitedto, cetyl alcohol, glycerol monostearate, polyoxyethylene sorbitan fattyacid esters (e.g., the commercially available Tweens® such as e.g.,Tween 20® and Tween 80® (ICI Specialty Chemicals)), and polyethyleneglycols (e.g., Carbowaxs 3350® and 1450®, and Carbopol 934® (UnionCarbide)), oleic acid, glyceryl monostearate, sorbitan monooleate,sorbitan monolaurate, triethanolamine oleate, polyoxyethylene sorbitanmonooleate, polyoxyethylene sorbitan monolaurate, sodium oleate, sodiumlauryl sulfate, sodium docusate, triacetin, vitamin E TPGS, sodiumtaurocholate, simethicone, phosphotidylcholine and the like

Suitable preservatives for the aqueous suspensions or dispersionsdescribed herein include, for example, potassium sorbate, parabens(e.g., methylparaben and propylparaben), benzoic acid and its salts,other esters of parahydroxybenzoic acid such as butylparaben, alcoholssuch as ethyl alcohol or benzyl alcohol, phenolic compounds such asphenol, or quaternary compounds such as benzalkonium chloride.Preservatives, as used herein, are incorporated into the dosage form ata concentration sufficient to inhibit microbial growth.

Suitable viscosity enhancing agents for the aqueous suspensions ordispersions described herein include, but are not limited to, methylcellulose, xanthan gum, carboxymethyl cellulose, hydroxypropylcellulose, hydroxypropylmethyl cellulose, Plasdon® S-630, carbomer,polyvinyl alcohol, alginates, acacia, chitosans and combinationsthereof. The concentration of the viscosity enhancing agent will dependupon the agent selected and the viscosity desired.

In addition to the additives listed above, the liquid formulations canalso include inert diluents commonly used in the art, such as water orother solvents, solubilizing agents, and emulsifiers. Exemplaryemulsifiers are ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethylacetate, benzyl alcohol, benzyl benzoate, propyleneglycol,1,3-butyleneglycol, dimethylformamide, sodium lauryl sulfate, sodiumdoccusate, cholesterol, cholesterol esters, taurocholic acid,phosphotidylcholine, oils, such as cottonseed oil, groundnut oil, corngerm oil, olive oil, castor oil, and sesame oil, glycerol,tetrahydrofurfuryl alcohol, polyethylene glycols, fatty acid esters ofsorbitan, or mixtures of these substances, and the like.

In some embodiments, the pharmaceutical compositions (e.g., HC-HA, fetalsupport tissue extracts, fetal support powders and homogenates)described herein can be self-emulsifying drug delivery systems (SEDDS).Emulsions are dispersions of one immiscible phase in another, usually inthe form of droplets. Generally, emulsions are created by vigorousmechanical dispersion. SEDDS, as opposed to emulsions or microemulsions,spontaneously form emulsions when added to an excess of water withoutany external mechanical dispersion or agitation. An advantage of SEDDSis that only gentle mixing is required to distribute the dropletsthroughout the solution. Additionally, water or the aqueous phase can beadded just prior to administration, which ensures stability of anunstable or hydrophobic active ingredient. Thus, the SEDDS provides aneffective delivery system for oral and parenteral delivery ofhydrophobic active ingredients. SEDDS may provide improvements in thebioavailability of hydrophobic active ingredients. Methods of producingself-emulsifying dosage forms are known in the art and include, but arenot limited to, for example, U.S. Pat. Nos. 5,858,401, 6,667,048, and6,960,563, each of which is specifically incorporated by reference.

Gels

In some embodiments, the compositions (e.g., HC-HA, fetal support tissueextracts, fetal support powders and homogenates) disclosed herein areformulated as a gel. In some embodiments, the compositions (e.g., HC-HA,fetal support tissue extracts, fetal support powders and homogenates)disclosed herein are formulated as solvent release gels. In someembodiments, the compositions (e.g., HC-HA, fetal support tissueextracts, fetal support powders and homogenates) disclosed herein areformulated as actinic radiation curable gels. In some embodiments, thecompositions (e.g., HC-HA, fetal support tissue extracts, fetal supportpowders and homogenates) disclosed herein are formulated asthermoreversible gels. In other embodiments, the compositions (e.g.,HC-HA, fetal support tissue extracts, fetal support powders andhomogenates) disclosed herein are formulated hydrogels.

As used herein, “gels”, sometimes referred to as jellies, are semisolidsystems consisting of either suspensions made up of small inorganicparticles or large organic molecules interpenetrated by a liquid. Gelsinclude a single-phase or a two-phase system. A single-phase gelconsists of organic macromolecules distributed uniformly throughout aliquid in such a manner that no apparent boundaries exist between thedispersed macromolecules and the liquid. Some single-phase gels areprepared from synthetic macromolecules (e.g., carbomer) or from naturalgums, (e.g., tragacanth). In some embodiments, single-phase gels aregenerally aqueous, but will also be made using alcohols and oils.Two-phase gels consist of a network of small discrete particles.

Gels are hydrophobic or hydrophilic. The base of a hydrophobic gel maybe liquid paraffin with polyethylene or fatty oils gelled with colloidalsilica, or aluminum or zinc soaps. The base of hydrophobic gels usuallyconsists of water, glycerol, or propylene glycol gelled with a suitablegelling agent (e.g., tragacanth, starch, cellulose derivatives,carboxyvinylpolymers, and magnesium-aluminum silicates).

Polymers composed of polyoxypropylene and polyoxyethylene formthermoreversible gels when incorporated into aqueous solutions. Thesepolymers have the ability to change from the liquid state to the gelstate at temperatures close to body temperature.

Paints

In some embodiments, the compositions (e.g., HC-HA, fetal support tissueextracts, fetal support powders and homogenates) disclosed herein areformulated as paints. As used herein, a “paint”, also known as filmformers, means solutions comprised of a solvent, a monomer or polymer,an tissue product or extract thereof, and optionally one or morepharmaceutically-acceptable excipients. After application to a targetbone, the solvent evaporates leaving behind a thin coating comprised ofthe monomers or polymers, and the active agent. The coating protectsactive agents and maintains them in an immobilized state at the site ofapplication. This decreases the amount of active agent that may be lostand correspondingly increases the amount delivered to the subject. Byway of non-limiting example, paints include collodions, and solutionscomprising saccharide siloxane copolymers and a cross-linking agent.Collodions are ethyl ether/ethanol solutions containing pyroxylin (anitrocellulose). After application, the ethyl ether/ethanol solutionevaporates leaving behind a thin film of pyroxylin. In solutionscomprising saccharide siloxane copolymers, the saccharide siloxanecopolymers form the coating after evaporation of the solvent initiatesthe cross-linking of the saccharide siloxane copolymers. The paintscontemplated for use herein, are flexible such that they do notinterfere with the growth or movement of a bone or joint. In someembodiments, the paints are applied as a liquid (i.e. solution,suspension, or emulsion), a semisolid (i.e. a gel, foam, paste, orjelly) or an aerosol.

Foams

In some embodiments, the compositions (e.g., HC-HA, fetal support tissueextracts, fetal support powders and homogenates) disclosed herein isformulated as a foam. Examples of suitable foamable carriers for use inthe compositions e.g., HC-HA, fetal support tissue extracts, fetalsupport powders and homogenates) disclosed herein include, but are notlimited to, alginate and derivatives thereof, carboxymethylcellulose andderivatives thereof, collagen, polysaccharides, including, for example,dextran, dextran derivatives, pectin, starch, modified starches such asstarches having additional carboxyl and/or carboxamide groups and/orhaving hydrophilic side-chains, cellulose and derivatives thereof, agarand derivatives thereof, such as agar stabilized with polyacrylamide,polyethylene oxides, glycol methacrylates, gelatin, gums such asxanthum, guar, karaya, gellan, arabic, tragacanth and locust bean gum,salts thereof (e.g., sodium alginate), or combinations thereof.

Liposomes

In some embodiments, the compositions (e.g., HC-HA, fetal support tissueextracts, fetal support powders and homogenates) disclosed herein areformulated as liposomes or lipid particles. Phospholipids that aregently dispersed in an aqueous medium form multilayer vesicles withareas of entrapped aqueous media separating the lipid layers.Sonication, or turbulent agitation, of these multilayer vesicles resultsin the formation of single layer vesicles, commonly referred to asliposomes. Suitable phospholipids for use in the formation of liposomes,include for example, phosphatidyl cholines, ethanolamines and serines,sphingomyelins, cardiolipins, plasmalogens, phosphatidic acids andcerebrosides. Preferred phospholipids are, for example, phosphatidylcholine, phosphatidyl ethanolmine, phosphatidyl serine, phosphatidylinositol, lysophosphatidyl choline, phosphatidyl glycerol.

In some embodiments, a liposome formulation may further comprise alipophilic additive. Examples of such additives include by way ofexample only, stearylamine, phosphatidic acid, tocopherol, cholesterol,cholesterol hemisuccinate and lanolin extracts.

Creams and Lotions

In some embodiments, the compositions (e.g., HC-HA, fetal support tissueextracts, fetal support powders and homogenates) disclosed herein areformulated as creams. In certain instances, creams are semisolid (e.g.,soft solid or thick liquid) formulations that include a fetal supporttissue product described herein (e.g., a flat tissue product sheet, apulverized tissue product, or a homogenized tissue product) dispersed inan oil-in-water emulsion or a water-in-oil emulsion.

Ointments

In some embodiments, the compositions (e.g., HC-HA, fetal support tissueextracts, fetal support powders and homogenates) disclosed herein areformulated as ointments. In certain instances, ointments are semisolidpreparations that soften or melt at body temperature.

Pastes

In some embodiments, the compositions (e.g., HC-HA, fetal support tissueextracts, fetal support powders and homogenates) disclosed herein areformulated as pastes. In certain instances, pastes contain at least 20%solids. In certain instances, pastes are ointments that do not flow atbody temperature.

Patches

In some embodiments, the compositions (e.g., HC-HA, fetal support tissueextracts, fetal support powders and homogenates) disclosed herein areadministered via a patch. In some embodiments, the compositionsdescribed herein are dissolved and/or dispersed in a polymer or anadhesive. In some embodiments, a film, a patch disclosed herein isconstructed for continuous, pulsatile, or on demand delivery of acomposition described herein.

Wound Dressings

In some embodiments, the compositions (e.g., HC-HA, fetal support tissueextracts, fetal support powders and homogenates) disclosed herein areadministered via a wound dressing. Wound dressings include, but are notlimited to gauzes, transparent film dressings, hydrogels, polyurethanefoam dressings, hydrocolloids and alginates.

Implants/Prosthesis

In some embodiments, the compositions (e.g., HC-HA, fetal support tissueextracts, fetal support powders and homogenates) disclosed herein areadministered via implants and prostheses. In some embodiments, thecompositions (e.g., HC-HA, fetal support tissue extracts, fetal supportpowders and homogenates) are administered by bone implants or bonestents.

In some embodiments, the compositions (e.g., HC-HA, fetal support tissueextracts, fetal support powders and homogenates) disclosed herein areadministered via a bone implant. In some embodiments, the bone implantis an osseointegrated implant. As used herein, an “osseointegratedimplant” means a three dimensional implant containing pores into whichosteoblasts and supporting connective tissue can migrate. In someembodiments, the bone implant comprises a composition described herein.In some embodiments, the bone implant is a dental implant. In someembodiments, the bone implant is used for knee or joint replacement. Insome embodiments, the bone implant is a craniofacial prosthesis (e.g.,an artificial ear, orbital prosthesis, nose prosthesis).

In some embodiments, the compositions (e.g., HC-HA, fetal support tissueextracts, fetal support powders and homogenates) disclosed herein areadministered via a bone stent. In some embodiments, the compositions(e.g., HC-HA, fetal support tissue extracts, fetal support powders andhomogenates) disclosed herein are coated onto the outside of the stent.In some embodiments, the compositions (e.g., HC-HA, fetal support tissueextracts, fetal support powders and homogenates) disclosed herein elutefrom the stent into the surrounding bone. In some embodiments, the bonestents are inserted into the intramedullary canal of a bone. In someembodiments, the bone stent is placed in the sinus tarsi. In someembodiments, the bone stent in placed in a knee or joint. In someembodiments, the bone stent is placed in a bone fracture. In someembodiments, the bone stent is expandable or contractable.

Miscellaneous

In some embodiments, the pharmaceutical composition described hereincomprise microencapsulated tissue products and/or extracts thereof(e.g., HC-HA). In some embodiments, one or more other compatiblematerials are present in the microencapsulation material. Exemplarymaterials include, but are not limited to, pH modifiers, erosionfacilitators, antifoaming agents, antioxidants, flavoring agents, andcarrier materials such as binders, suspending agents, disintegrationagents, filling agents, surfactants, solubilizers, stabilizers,lubricants, wetting agents, and diluents.

Exemplary microencapsulation materials useful for delaying the releaseof the formulations including compounds described herein, include, butare not limited to, hydroxypropyl cellulose ethers (HPC) such as Klucel®or Nisso HPC, low-substituted hydroxypropyl cellulose ethers (L-HPC),hydroxypropyl methyl cellulose ethers (HPMC) such as Seppifilm-LC,Pharmacoat®, Metolose SR, Methocel®-E, Opadry YS, PrimaFlo, BenecelMP824, and Benecel MP843, methylcellulose polymers such as Methocel®-A,hydroxypropylmethylcellulose acetate stearate Aqoat (HF-LS, HF-LG,HF-MS) and Metolose®, Ethylcelluloses (EC) and mixtures thereof such asE461, Ethocel®, Aqualon®-EC, Surelease®, Polyvinyl alcohol (PVA) such asOpadry AMB, hydroxyethylcelluloses such as Natrosol®,carboxymethylcelluloses and salts of carboxymethylcelluloses (CMC) suchas Aqualon®-CMC, polyvinyl alcohol and polyethylene glycol co-polymerssuch as Kollicoat IR®, monoglycerides (Myverol), triglycerides (KLX),polyethylene glycols, modified food starch, acrylic polymers andmixtures of acrylic polymers with cellulose ethers such as Eudragit®EPO, Eudragit® L30D-55, Eudragit® FS 30D Eudragit® L100-55, Eudragit®L100, Eudragit® S100, Eudragit® RD100, Eudragit® E100, Eudragit® L12.5,Eudragit® S12.5, Eudragit® NE30D, and Eudragit® NE 40D, celluloseacetate phthalate, sepifilms such as mixtures of HPMC and stearic acid,cyclodextrins, and mixtures of these materials.

In still other embodiments, plasticizers such as polyethylene glycols,e.g., PEG 300, PEG 400, PEG 600, PEG 1450, PEG 3350, and PEG 800,stearic acid, propylene glycol, oleic acid, and triacetin areincorporated into the microencapsulation material. In other embodiments,the microencapsulating material useful for delaying the release of thepharmaceutical compositions is from the USP or the National Formulary(NF). In yet other embodiments, the microencapsulation material isKlucel. In still other embodiments, the microencapsulation material ismethocel.

In some embodiments, the fetal support tissue products or extractsthereof (e.g., HC-HA) are incorporated within nanoparticles,microparticles, or microspheres.

Microspheres usually have a spherical shape, although irregularly-shapedmicroparticles are possible. Microspheres may vary in size, ranging fromsubmicron to 1000 micron diameters. The fetal support tissue productsand extracts thereof are encapsulated in microspheres by any suitablemethods. Generally, the agent is dispersed or emulsified, usingstirrers, agitators, or other dynamic mixing techniques, in a solventcontaining a wall-forming material. Solvent is then removed from themicrospheres, and thereafter the microsphere product is obtained.

Nanoparticles are also contemplated for use with the fetal supporttissue products and extracts described herein. Nanoparticles arematerial structures of about 100 nm or less in size. One use ofnanoparticles in pharmaceutical compositions is the formation ofsuspensions as the interaction of the particle surface with solvent isstrong enough to overcome differences in density. Nanoparticlesuspensions are sterilized as the nanoparticles are small enough to besubjected to sterilizing filtration (see, e.g., U.S. Pat. No. 6,139,870,herein incorporated by reference for such disclosure). Nanoparticlescomprise at least one hydrophobic, water-insoluble andwater-indispersible polymer or copolymer emulsified in a solution oraqueous dispersion of surfactants, phospholipids or fatty acids. Thenanoparticles may be obtained by any suitable methods. These methodsinclude vaporization methods, such as free jet expansion, laservaporization, spark erosion, electro explosion and chemical vapordeposition; physical methods involving mechanical attrition (e.g.,“pearlmilling” technology, Elan Nanosystems), super critical CO2 andinterfacial deposition following solvent displacement. In oneembodiment, the solvent displacement method is used. The size ofnanoparticles produced by this method is sensitive to the concentrationof polymer in the organic solvent; the rate of mixing; and to thesurfactant employed in the process. Continuous flow mixers provide thenecessary turbulence to ensure small particle size. One type ofcontinuous flow mixing device that is optionally used to preparenanoparticles has been described (Hansen et al J Phys Chem 92, 2189-96,1988). In other embodiments, ultrasonic devices, flow throughhomogenizers or supercritical CO2 devices may be used to preparenanoparticles.

Combination Therapies

Disclosed herein, in certain embodiments, are methods of inhibitingosteoclast differentiation in an individual in need thereof, comprisingadministering to the individual a composition comprising a fetal supporttissue product or an extract thereof. Disclosed herein, in certainembodiments, are methods of inhibiting bone resorption in an individualin need thereof, comprising administering to the individual acomposition comprising a fetal support tissue product or an extractthereof. Disclosed herein, in certain embodiments, are methods ofinhibiting bone remodeling in an individual in need thereof, comprisingadministering to the individual a composition comprising a fetal supporttissue product or an extract thereof. Disclosed herein, in certainembodiments, are methods of balancing bone resorption and boneformation, comprising administering to an individual in need thereof atherapeutically-effective amount of a composition comprising a fetalsupport tissue product or an extract thereof. Disclosed herein, incertain embodiments, are methods of treating a disease, disorder, orcondition characterized by excessive or undesired osteoclastdifferentiation, the methods comprising administering to an individualin need thereof a therapeutically-effective amount of a compositioncomprising a fetal support tissue product or an extract thereof.Disclosed herein, in certain embodiments, are methods of treatingarthritis, the methods comprising administering to an individual in needthereof a therapeutically-effective amount of a composition comprising afetal support tissue product or an extract thereof. In some embodiments,the arthritis is osteoarthritis, rheumatoid arthritis, psoriaticarthritis, or any combination thereof. Disclosed herein, in certainembodiments, are methods of treating osteoporosis, the methodscomprising administering to an individual in need thereof atherapeutically-effective amount of a composition comprising a fetalsupport tissue product or an extract thereof. Disclosed herein, incertain embodiments, are methods of treating alveolar bone degradation,the methods comprising administering to an individual in need thereof atherapeutically-effective amount of a composition comprising a fetalsupport tissue product or an extract thereof. Disclosed herein, incertain embodiments, are methods of treating Paget's disease, themethods comprising administering to an individual in need thereof atherapeutically-effective amount of a composition comprising a fetalsupport tissue product or an extract thereof. Disclosed herein, incertain embodiments, are methods of treating a bone tumor, the methodscomprising administering to an individual in need thereof atherapeutically-effective amount of a composition comprising a fetalsupport tissue product or an extract thereof. In some embodiments of anyof the aforementioned methods, the methods further compriseadministering a further therapeutic agent.

In some embodiments, the pharmaceutical compositions disclosed hereinresult from the mixing or combining of more than one active ingredientand includes both fixed and non-fixed combinations of the activeingredients. The term “fixed combination” means that the activeingredients, e.g. a compound described herein and a co-agent, are bothadministered to a patient simultaneously in the form of a single entityor dosage. The term “non-fixed combination” means that the activeingredients, e.g. a compound described herein and a co-agent, areadministered to a patient as separate entities either simultaneously,concurrently or sequentially with no specific intervening time limits,wherein such administration provides effective levels of the twocompounds in the body of the patient. The latter also applies tococktail therapy, e.g. the administration of three or more activeingredients.

In some embodiments, the further therapeutic agent is a calciumsupplement, a calcium salt, or a combination thereof. In someembodiments, the further therapeutic agent is calcium carbonate, coralcalcium, calcium citrate, calcium phosphate, calcium lactate, a calciumchelate (e.g., calcium malate, calcium aspartate, calcium fumarate), orany combinations thereof.

In some embodiments, the further therapeutic agent is a NSAID. In someembodiments, the further therapeutic agent is a salicylate, a propionicacid derivative, an acetic acid derivative, and enolic acid derivative,a fenamic acid derivative, a selective COX-2 inhibitor, asulphonanilide, or any combinations thereof. In some embodiments, thefurther agent is aspirin, diflunisal, salsalate, ibuprofen, naproxen,fenoprofen, ketoprofen, dexketoprofen, flurbiprofen, oxaprozin,loxoprofen, indomethacin, sulindac, etodolac, ketorolac, diclofenac,nabumetone, piroxicam, meloxicam, tenoxicam, droxicam, lornoxicam,isoxicam, mefenamic acid, meclofenamic acid, flufenamic acid, tolfenamicacid, celecoxib, rofecoxib, valdecoxib, parecoxib, lumiracoxib,etoricoxib, firocoxibm, nimesulide, licofelone, or any combinationsthereof.

In some embodiments, the further therapeutic agent is a corticosteroid.In some embodiments, the further therapeutic agent is a hydrocortisonetype corticosteroid, an acetonide, a betamethasone type corticosteroid,a corticosteroid ester (e.g., a halogenated corticosteroid or a labileprodrug ester). In some embodiments, the further therapeutic agent ishydrocortisone, hydrocortisone acetate, cortisone acetate, tixocortolpivalate, prednisolone, methylprednisolone, prednisone, triamcinoloneacetonide, triamcinolone alcohol, mometasone, amcinonide, budesonide,desonide, fluocinonide, fluocinolone acetonide, halcinonide,betamethasone, betamethasone sodium phosphate, dexamethasone,dexamethasone sodium phosphate, fluocortolone,hydrocortisone-17-valerate, aclometasone dipropionate, betamethasonevalerate, betamethasone dipropionate, prednicarbate,clobetasone-17-butyrate, clobetasol-17-propionate, fluocortolonecaproate, fluocortolone pivalate, and fluprednidene acetate,hydrocortisone-17-butyrate, 17-aceponate, 17-buteprate, Prednicarbate,or any combinations thereof.

In some embodiments, the further therapeutic agent is a hyaluronan orsodium hyaluronate.

In some embodiments, the further therapeutic agent is a DMARD(disease-modifying antirheumatic drug). In some embodiments, the furthertherapeutic agent is adalimumab, azathioprine, chloroquine,hydroxychloroquine, cyclosporine, D-penicillamine, etanerecpt,golimumab, infliximab, leeflunomide, methotrexatem minocycline,rituximab, sulfasalazine, sodium aurothiomalate, auranofin, or anycombinations thereof.

In some embodiments, the further therapeutic agent is a bisphosphonate.In some embodiments, the further therapeutic agent is a N-containingbisphosphonate. In some embodiments, the further therapeutic agent is anon-N-containing bisphosphonate. In some embodiments, the furthertherapeutic agent is etidronate, clodronate, tiludronate, pamidronate,neridronate, olpadronate, alendronate, ibandronate, risedronate,zoledronate, or any combination thereof.

In some embodiments, the further therapeutic agent is an estrogenanalog. In some embodiments, the further therapeutic agent is estrodiol,phytoestrogen, diethylstilbestrol, or any combinations thereof.

In some embodiments, the further therapeutic agent is a selectiveestrogen receptor modulator (SERM). In some embodiments, the furthertherapeutic agent is raloxifene, clomifene, femarelle, ormeloxifene,tamoxifen, toremifene, lasofoxifene, or any combinations thereof.

In some embodiments, the further therapeutic agent is a member of thecalcitonin-like protein family. In some embodiments, the furthertherapeutic agent is Calcitonin.

In some embodiments, the further therapeutic agent is parathyroidhormone. In some embodiments, the further therapeutic agent is arecombinant parathyroid hormone. In some embodiments, the furthertherapeutic agent is Teriparatide.

In some embodiments, the further therapeutic agent is a RANKL inhibitor.In some embodiments, the further therapeutic agent is denosumab.

In some embodiments, the further therapeutic agent is achemotherapeutic. Any suitable chemotherapeutic agent is contemplatedfor use with the fetal support tissue product compositions disclosedherein. Suitable chemotherapeutic agents include those that areefficacious in the treatment of osteosarcoma, Ewing's sarcoma,chondrosarcoma, and fibrosarcoma. In some embodiments, the furthertherapeutic agent is ifosfamide, etopside, carboplatin, cisplatin,cyclophosphamide, doxorubicin, epirubicin, methotrexate, PEG-interferonalfa-2b, or any combinations thereof.

In some embodiments, the further therapeutic agent is radiation therapy,proton therapy, or a combination thereof.

In some embodiments, the further therapeutic agent is sodium fluoride.

In some embodiments, the further therapeutic agent is Strontiumranelate. In some embodiments, the further therapeutic agent isMetastron (i.e., strontium-89 chloride).

EXAMPLES Example 1 AM Tissue and its Derivatives Prevent Differentiationof Osteoclasts in Response to RANKL

Cell Culture and Treatment

Macrophage RAW 264.7 cells were cultivated in DMEM/10% FBS until 80%confluence, harvested with enzyme-free dissociation buffer, and seededat 169 cells/mm² with α-MEM/10% FBS differentiation media without phenolred.

After 24 h, seeded macrophage RAW 264.7 cells were treated with 50 ng/mlRANKL (except Neg CTL) and simultaneously treated with either PBS 1X(Pos CTL), AME (200 μg/ml Protein), HC-HA (25 μg/ml HA), or on eitherside (epithelium, stroma or basement membrane) of intact AM (iAM),denuded AM (dAM), and lyophilized AM (lAM). Cells were observed for celldeath and formation of multinucleated cell (osteoclast) for a period of5-7 days.

TRAP Assay

After termination, cells were fixed with acetone and stained withtartrate resistant acid phosphatase leukocyte kit (TRAP). Cell lysatewas also collected with 1% Triton X-100 extraction buffer for analysiswith TRAP Colorimetric Assay.

Formation of large multinucleated osteoclast cells was observed for thePos CTL (FIG. 2 E-G) but not Neg CTL (FIG. 2 A-C). TRAP staining (FIG.3) revealed no large multinucleated osteoclast cells when RAW 264.7cells were cultured on the epithelial side of intact amniotic membrane(iAM), the stromal side of iAM (F), the basement membrane side ofepithelially-denuded AM (dAM) (G), the stromal side of dAM (H), theepithelial side of lyophilized amniotic membrane lAM (I), or the stromalside of lyophilized amniotic membrane lAM (J).

Following TRAP Colorimeteric analysis, the Neg CTL showed significantlylower (p=2.9E-12) TRAP reading compared to the Pos CTL. Inhibitoryaction for osteoclast formation was seen for all AM derivatives (FIG. 4)with or without gamma irradiation.

qPCR

Total RNAs were also collected for quantitative measurement of RANK,β3-Integrin, and NFATc1 transcripts.

Quantitative RT-PCR revealed that levels of NFATc1 mRNA weresignificantly downregulated (p<0.05) in response to all AM derivatives(FIG. 5). Expression of RANK was found to be significantly downregulatedfor both HC-HA and stromal side of dAM (FIG. 7). Expression ofβ3-integrin (FIG. 8), which is required for normal osteoclastdifferentiation, was found to be significantly downregulated by HC-HA,AML, AMP, and stromal side of dAM. Suppression of β3-integrin by AME isstatistically significant on Day 5 (p=0.000001).

Example 2 AME, AML, and AMP Inhibit Osteoclast Formation from RANKLStimulated RAW 264.7 Macrophage Cells

Murine RAW 264.7 macrophage cells were seeded at a density of 4.0×10³cells/96 well (30-40% confluent), cultured in α-MEM media without PhenolRed and supplemented with 10% FBS, 100 μg/ml penicillin & streptomycin.24 hours after seeding, cells were treated with or without 50 ng/mlRANKL stimulation. Experimental groups were simultaneously treated withAMP, AML or AME with protein concentration of 200 μg/ml. On Day 5, theculture was terminated and analyzed by TRAP staining and TRAPColorimetric Assay.

The result from TRAP staining (FIG. 2) shows that osteoclasts(multi-nucleated cells) were not found on the Neg CTL while largemulti-nucleated cells were found on the Pos CTL. AME inhibitedosteoclast formation but did not inhibit RAW macrophage cellproliferation (FIG. 2a ). Osteoclast formation and RAW macrophage cellproliferation were also inhibited by AML-1 and AML-2 from 2 differentdonors (FIG. 11), but such inhibition was not complete because smallmulti-nucleated cells could be seen after TRAP staining Osteoclastformation and RAW macrophage cell proliferation were also inhibited byAMP from 5 different donors (FIG. 11) at the same protein concentrationof 200 μg/ml as AML.

The result from TRAP staining shows that the inhibitory action forosteoclast formation was seen on all AM derivatives. There was nosignificant difference between the TRAP Colorimetric Assay readingbetween AMP and AME (Table 1). For two donors, the TRAP ELISA readingfor AMP is significantly lower than AML (Table 1) from the same donor(p=0.04 and p=0.02) at the same protein concentration 200 μg protein/ml(FIG. 11).

TABLE 1 p-values Comparison p = value AMP 1-5 AME 0.24 AMP 1-2 AML 1-21.02E−05

TABLE 2 OD readings and p-values OD ± standard p-value Conditionsdeviation (compared to Pos CTL) Neg CTL 0.037 ± 0.0035 0.0013 Pos CTL 0.53 ± 0.11 — AME 0.073 ± 0.015 0.0028 HC.HA (25 μg/ml HA) 0.023 ±0.0081 0.0023 AML-1  0.18 ± 0.0097 0.0067 AMP-1 0.046 ± 0.018 0.0022AML-2  0.19 ± 0.052 0.0034 AMP-2 0.093 ± 0.0055 0.0036 Mean AMP 1-50.087 ± 0.041 0.0017

TABLE 3 Protein Content of AMP from 5 different donors BCA ProteinPowder Protein (μg)/ Total Total Protein Reading used/ml Powder WeightPowder (mg) from (μg/ml) (mg/ml) (mg) Weight (mg) 1 whole AM AMP-1 1393344.6 312 1340 419 AMP-2 8777 30 293 880 257 AMP-3 14527 42 346 1250 432AMP-4 16516 49 337 1836 619 AMP-5 22727 51.2 444 1408 625

Example 3 Powder Derived from Amniotic Membrane (AMP), Chorion (CHP),Amnio-Chorion (ACP), Placenta (PCP), Whole Umbilical Cord (UCP), orUmbilical Cord AM (UCAP) Inhibits Osteoclast Formation from RANKLStimulated RAW 264.7 Macrophage Cells

Murine RAW 264.7 macrophage cells were seeded at a density of 4.0×10³cells/96 well (30-40% confluent), cultured in α-MEM media without PhenolRed and supplemented with 10% FBS, 100 μg/ml penicillin & streptomycin.24 hours after seeding, cells were treated with or without 50 ng/mlRANKL stimulation. Experimental groups were simultaneously treated withAMP, CHP, ACP, PLP, UCP, and UCAP with protein concentration of 100μg/ml. On Day 5, the culture was terminated and analyzed by TRAPColorimetric Assay.

Osteoclast formation was inhibited by all powder derived from amnioticmembrane (AMP), chorion (CHP), amnio-chorion (ACP), placenta (PCP),whole umbilical cord (UCP), and umbilical cord AM (UCAP) (FIG. 12).

Example 4 HC-HA Inhibits the Formation of Osteoclasts from RANKLStimulated RAW 264.7 Macrophage Cells

Murine RAW 264.7 macrophage cells were seeded at a density of 4.0×10³cells/96 well (30-40% confluent), cultured in α-MEM media without PhenolRed and supplemented with 10% FBS, 100 μg/ml penicillin & streptomycin.24 hours after seeding, cells were treated with or without 50 ng/mlRANKL stimulation.

Experimental groups were simultaneously treated with high molecularweight HA or HC-HA prepared at a series of HA concentrations (0, 0.008,0.04, 0.2, 1, 5, and 25 μg/ml). On Day 5, the culture was terminated andanalyzed by TRAP staining and TRAP ELISA.

The result from TRAP staining shows that osteoclasts (multi-nucleatedcells) are not found on the negative control while large multi-nucleatedcells are found on the positive control (FIG. 9a ).

Even at the highest concentration (25 μg/ml), HA does not inhibits theformation of multinucleated cells and the expression of TRAP, Incontrast, the inhibition by HC-HA is detected as low as at 0.08 μg/ml,and dose-dependently increased until the multinucleated cells arecompletely eliminated at 25 μg/ml (FIG. 9 b).

These observations are also confirmed by quantitatively measured TRAPactivity (FIGS. 9c and d ). From the TRAP Colorimetric assay, the IC50of HC-HA's inhibition on the osteoclast formation induced by RANKL is0.1 μg/ml (FIG. 9e ).

Example 5 AMP Promotes the Mineralization of Osteoblasts

Osteoblast precursor MC3T3-E1 cells were maintained in DMEM/10% FBS butwere re-suspended into α-MEM/10% FBS and seeded at 1×10⁵/ml on 24 wellplastic (2 ml per well, and designated as Day 1 from here onwards) for 2days.

The culture medium was replaced with either PBS (Neg. Ctrl),osteoblast-inducing reagents (0.2 mM ascorbic acid 2-phosphate and 10 mMglycerol 2-phosphate, Pos. Ctrl), or osteoblast-inducing reagents plus0.1 μg/ml HC-HA or 125 μg/ml AMP. The cell culture medium was changedevery 3 days until on Day 18. From Day 8 onwards, theosteoblast-inducing reagents was additionally supplemented withmelatonin (50 ng/ml).

At Day 18, cells were assayed with alizarin red staining (ARS) tomeasure the mineralization of differentiated osteoblasts. Negativecontrol was not stained by ARS but the positive control was stained inred (FIG. 13).

Treatment with HC-HA (0.1 μg/ml) has a similar staining as that of thepositive control (FIG. 13). However, treatment with AMP (125 μg/ml)yields much darker staining (AMP), indicating that more minerals aregenerated (FIG. 14).

The differences were also observed after ARS stained cells were removedby 4 M GnCl, showing less cells were left in AMP treated wells (FIG.14). Some cells migrate from the monolayer into the AMP particles anduse it as a scaffold for differentiation and mineralization.

Quantitative measurement of ARS (FIG. 14) shows HC-HA (0.1 μg/ml) doesnot significantly increase or decrease the mineralization when comparedto the positive control (p>0.05). In contrast, cells treated with AMP(125 μg/ml) significantly promotes the mineralization compared to eitherthe positive control or HC-HA (p=0.0001). The color changes related tothe concentrations of alizarin red or in samples are shown in FIG. 14.

Example 6 Use for the Treatment of an Arthritic Joint

An individual with rheumatoid arthritis is identified. A pharmaceuticalcomposition for injection comprising HC-HA is prepared.

The composition is injected at the arthritic joints of the individual.

Example 7 Use in the Treatment of Osteolysis

An individual with an area of decalcified bone is identified. A wounddressing comprising placental powder is prepared.

The tubular tissue graft is surgically placed around the area ofdecalcified bone.

Example 8 Use of an Implant in the Treatment of Osteolysis

An individual with an area of decalcified bone is identified. An implantcoated in HC-HA is prepared.

The implant is surgically placed near the area of decalcified bone.

Example 9 Use of an Patch in the Treatment of Alveolar Bone Degradation

An individual with alveolar bone degradation is identified. An sheet ofsubstantially-flat amnion-chorion patch is prepared.

The patch is placed on an alveolar bone at the site of alveolar bonedegradation.

Example 10 Use of an Orthopaedic Prosthesis in the Treatment ofOsteoarthritis

An individual with a damaged knee due to osteoarthritis is identified.An orthopedic prosthesis comprising chorion homogenate derivative isprepared.

The orthopedic prosthesis is surgically placed to replace theindividual's damaged knee.

While preferred embodiments of the present invention have been shown anddescribed herein, it will be obvious to those skilled in the art thatsuch embodiments are provided by way of example only. Numerousvariations, changes, and substitutions will now occur to those skilledin the art without departing from the invention. It should be understoodthat various alternatives to the embodiments of the invention describedherein may be employed in practicing the invention. It is intended thatthe following claims define the scope of the invention and that methodsand structures within the scope of these claims and their equivalents becovered thereby.

What is claimed is:
 1. A method of inhibiting unwanted bone resorptionin an individual in need thereof, comprising contacting a bone or jointcharacterized by unwanted bone resorption in said individual with atherapeutically-effective amount of a fetal support tissue thatcomprises: (a) cells, substantially all of which have been killed, and(b) HC-HA, thereby inhibiting unwanted bone resorption.
 2. The method ofclaim 1, wherein the fetal support tissue is placental amniotic membrane(PAM), umbilical cord amniotic membrane (UCAM), placenta, umbilicalcord, chorion, amnion-chorion, or any combinations thereof.
 3. Themethod of claim 1, wherein the fetal support tissue is frozen orpreviously frozen.
 4. The method of claim 1, wherein the fetal supporttissue is a sheet, powder, or homogenate.
 5. The method of claim 1,wherein the fetal support tissue is cryopreserved, lyophilized,terminally sterilized, or a combination thereof.
 6. The method of claim1, wherein contacting a joint characterized by unwanted bone resorptionwith the fetal support tissue comprises injecting the fetal supporttissue into the joint.
 7. The method of claim 1, wherein contacting abone or joint characterized by unwanted bone resorption with the fetalsupport tissue comprises contacting the bone or joint with a medicaldevice comprising the fetal support tissue.
 8. The method of claim 1,wherein the medical device is a patch, implant, orthopaedic prosthesis,or bone stent.
 9. The method of claim 1, wherein contacting a bone orjoint characterized by unwanted bone resorption with the fetal supporttissue comprises wrapping a fetal support tissue sheet around the boneor joint.
 10. The method of claim 1, wherein osteogenesis is promoted.11. The method of claim 1, wherein the unwanted bone resorption isassociated with arthritis, osteoporosis, a bone tumor, Paget's Disease,alveolar bone degradation, or any combination thereof.